HHV-8 Virus Research Articles

Immunophenotyping and viral studies in pityriasis lichenoides et varioliformis acuta lesions.

The underlying pathogenesis of pityriasis lichenoides et varioliformis acuta (PLEVA) remains unclear, although immunologic injury and viral etiology have been suggested. To evaluate and expand the immunophenotype of PLEVA and to search for possible viral pathogens. Formalin-fixed, paraffin-embedded specimens of 20 patients with PLEVA and 9 patients with common inflammatory dermatoses (ID) were studied for immunophenotyping and for human herpesvirus (HHV) 1 and 2, cytomegalovirus (CMV), HHV-8, parvovirus B19, and Epstein-Barr virus (EBV) immunohistochemistry. The presence of HHV-6, HHV-7, and enteroviruses was assayed molecularly. The numbers of CD8+ T cells and T-cell intracellular antigen-1 (TIA-1)+ cells were statistically significantly higher in PLEVA compared to the ID group. Immunohistochemistry for human HHV-1 and HHV-2, CMV and HHV-8, parvovirus B19, and in situ hybridization for EBV were all negative. There was molecular evidence for HHV-7 in only one PLEVA case (5%). Molecular studies for HHV-6 and enterovirus involvement were negative in all the PLEVA specimens. The predominant T-cell infiltrate in PLEVA is dominated by CD8+ cells, and by increased numbers of TIA1+ cells, which may indicate a cytotoxic T-cell damage to the epidermis. Viral presence was not detected.

Viral myocarditis in combination with genetic cardiomyopathy as a cause of sudden death. An autopsy series

Sudden cardiac death (SCD) is a major public health issue worldwide. In the young (< 40 years of age), genetic cardiomyopathies and viral myocarditis, sometimes in combination, are the most frequent, but underestimated, causes of SCD. Molecular autopsy is essential for prevention. Several studies have shown an association between genetic cardiomyopathies and viral myocarditis, which is probably underestimated due to insufficient post-mortem investigations. We report on four autopsy cases illustrating the pathogenesis of these combined pathologies. In two cases, a genetic hypertrophic cardiomyopathy was diagnosed in combination with Herpes Virus Type 6 (HHV6) and/or Parvovirus-B19 (PVB19) in the heart. In the third case, autopsy revealed a dilated cardiomyopathy and virological analyses revealed acute myocarditis caused by three viruses: PVB19, HHV6 and Epstein-Barr virus. Genetic analyses revealed a mutation in the gene coding for desmin. The fourth case illustrated a channelopathy and a PVB19/HHV6 coinfection. Our four cases illustrate the highly probable deleterious role of cardiotropic viruses in the occurrence of SCD in subjects with genetic cardiomyopathies. We discuss the pathogenetic link between viral myocarditis and genetic cardiomyopathy. Molecular autopsy is essential in prevention of these SCD, and a close collaboration between cardiologists, pathologists, microbiologists and geneticians is mandatory.

Եզակի օջախային Կաստլեմանի հիվանդության ախտորոշման և բուժման ժամանակակից մոտեցումները

Castleman disease is a rare lymphoproliferative disorder associated with HHV-8 and HIV viruses. Currently, there are two types of Castleman disease based on the prevalence of affected lymph nodes: Unicentric Castleman disease and Multicentric Castleman disease. Unicentric Castleman disease involves a lesion of one lymph node or lymph nodes in the same group. The diagnosis is made by histological and immunohistochemical examination of the removed lymph node. Furthermore, it is essential to distinguish Unicentric Castleman disease from Multicentric Castleman disease, for which PET/CT screening is an excellent method. The treatment approach of Unicentric Castleman disease is the surgical excision of all affected lymph nodes, and in non-resectable and asymptomatic cases, control is prescribed. In symptomatic cases, immunosuppressive therapy (rituximab, steroids, anti-IL6 antibodies) should be considered, followed by surgery or radiotherapy.

The Molecular Prevalence of Varicella-Zoster Virus and Human Herpesvirus Types 6 and 7 in Sperm Samples of Infertile Men

Background and Aims: Human herpes virus types 6 and 7 (HHV-6, HHV-7) can remain latent in the urogenital tract epithelial cells. Varicella-Zoster virus (VZV) is among the Alphaherpesviruses infecting most people during childhood. This virus can remain latent in the ganglionic neurons. The latent infection can reactivate through weakness, suppression of the immune system, and aging. Therefore, this study investigated the molecular prevalence of HHV-6, HHV-7, and Varicella-Zoster virus in sperm samples from infertile men.&#x0D; Materials and Methods: Sperm samples from 82 infertile patients were collected from infertility centers in Tehran. Chromatin health levels and sperm DNA examinations were carried out using sperm DNA fragmentation index kits. Viral nucleic acids were extracted from the samples. The presence of VZV, HHV-6, and HHV-7 genomes was examined through the conventional polymerase chain reaction assay.&#x0D; Results: The average age was 37.3 ± 6.1 years. The mean sperm motility percentage was 33.6 ± 2%. Infertile men's mean DNA fragmentation index was 27.2 ± 1.2. This study shows a 7.3% and 4.9% prevalence for HHV-6 and VZV, respectively, and the absence of HHV-7 in sperm specimens.&#x0D; Conclusion: Although the infection rates of these herpesviruses seem relatively low, more comprehensive and multi-center studies are required to obtain more accurate results.

Post-Transplant Cyclophosphamide, Abatacept and Short Course of Tacrolimus (CAST) for Graft-Versus-Host Disease Prevention Following Haploidentical Peripheral Blood Stem Cell Transplantation

Background: Haploidentical blood and marrow transplantation (BMT) substantially expands the pool of potential hematopoietic stem cell donors, including for minority groups. However, the outcomes of haploidentical transplantation remain inferior to matched unrelated donor (MUD) transplants when a standard post-transplant cyclophosphamide (PTCy)-based graft versus host disease (GvHD) prophylaxis regimen is employed in both settings. To address this issue, we sought to develop a more effective GvHD prevention regimen in haploidentical transplantation. While abatacept (Aba) seems to decrease the incidence of acute GvHD, PTCy is particularly effective in the prevention of chronic GvHD. Therefore, we designed a regimen that combines the merits of both agents and examined whether such approach can allow early withdrawal of tacrolimus (Tac). Methods: We enrolled 46 adult patients with hematological malignancies receiving haploidentical transplantation from a relative in a prospective single center pilot phase Ib-II clinical trial. Patients received non-myeloablative, reduced-intensity or myeloablative preparative regimens followed by G-CSF mobilized peripheral blood transplants and a combination of PTCy (50mg/kg on days +3 and +4), Aba (10mg/kg on days +5, +14 and +28) and Tac at a dose adjusted based on blood level. Tac taper was initiated on day +60 and completed by day +90. After treating 30 patients, the study was amended to administer an additional dose of Aba on day +56 based on the noted timing of acute GvHD development and Aba terminal ½ life. Patients received standard supportive care by institutional practice including G-CSF and anti-microbial prophylaxis. Serologically CMV positive patients also received letermovir prophylaxis. Results: Patient characteristics are summarized in table 1. Importantly, 19 out of 46 (41.3%) patients were from racial or ethnic minorities. The data cutoff date was July 29th, 2022. Three patients were excluded from this analysis due to short follow-up. For the remaining patients, median follow-up was 8.9 months. Median time to neutrophil engraftment was 18 days (13-30). One patient died before achieving platelet (plt) engraftment and 2 patients have not yet achieved plt engraftment. For the remaining 40 patients, median time to plt engraftment was 29 days (16-52). All patients achieved full donor chimerism at the time of count recovery. Tac was successfully tapered off as planned in all but 5 patients. Cumulative incidence of acute GvHD grades II-IV, III-IV and IV with death as competing event was 12.8%, 5.1% and 0%, respectively. One-year (y) cumulative incidence of moderate to severe chronic GvHD was 17.1%. All cases of acute GvHD except 1 grade II acute GvHD were observed in the first 30 patients. Cases of chronic GvHD included 2 cases of overlap syndrome. There was no case of steroid-refractory acute GvHD or GvHD-related death. One-y cumulative incidence of relapse (non-relapse mortality as competing event) was 8.3% (95% CI 0%-17.1%). KM estimates of 1-year relapse-free survival (RFS), overall survival (OS) and GRFS (composite end point of grade III-IV acute GvHD-, chronic GvHD requiring systemic therapy- and relapse-free survival) were 91.7% (95% CI 83%-100%), 90.6% (95% CI 80.9%-100%) and 71.8% (95% CI 57.2%-90%), respectively. None of the safety stopping rules was triggered. There was 1 case of thrombotic microangiopathy due to Tac and 2 cases of sinusoidal occlusive disease, 1 requiring treatment with defibrotide; all 3 patients recovered. There were 2 cases of respiratory failure of undetermined etiology. One-y cumulative incidence of non-relapse mortality (relapse as competing event) was 2.6% (95% CI 0%-7.8%). The incidence of CMV and EBV reactivation was 39.5% and 2.3%, respectively. There was no case of adenovirus or HHV-6 virus reactivation. BK viruria occurred in 27.9% of patients with limited symptomatology (1 patient required treatment with cidofovir). Immune reconstitution results are summarized in figure 1. Conclusion: CAST is a new GvHD prevention regimen that seems safe and effective, yielding excellent outcomes. CAST also allows early discontinuation of Tac. Outcomes with longer follow-up will be presented at the meeting. If the results are confirmed, CAST might be a step forward toward closing an important health disparity gap in BMT, equalizing the outcomes of haploidentical transplantation and MUD transplants. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Clinical Outcome of Unrelated Cord Blood Engraftment for Children with Juvenile Myelomonocytic Leukemia

Objective: To evaluate the clinical efficacy and safety of unrelated cord blood engraftment in the treatment of Juvenile Myelomonocytic Leukemia. Method: Seventy-three JMML children received cord blood transplantation since 2018. 47 children with cord blood engrafment were analyzed retrospectively. The median at diagnosis age was 35 months (12 to 99). Of them, 8 patients with NF-1 mutation, 24 with PTPN11 mutation, 2 with Nras somatic mutation, 4 with Kras somatic mutation, 8 with multiple mutation. The transplantation regimen was divided into two groups: Complementary transplantation group (CT-group) i.e. 55 patients underwent unrelated cord blood(UCB) following haploidentical stem cells transplantation(see blood 2016 128:1235 and blood 2019 134: 4181).UCB-cells engrafted finally in 31 patients,(others with haplo-engrafted).Conditioning regimen composed of Cyclophosphamide (Cy), Busulfan (Bu), Thiotepa (TT) and Fludarabine (Flu). Haploid PBSC (MNC >15×108/kg) was infused on day 0. GVHD prophylaxis consisted of PTCy on day+3 to +4,and FK506 and Mycophenolate Mofetil (MMF). Unrelated cord blood was infused on day+6. The next group was haploid DLI Bridging unrelated cord blood transplantation group (LCB-group) . Conditioning regimen composed of Cla+Ara-C for 4 days, and the haploid donor DLI was infused on day-11 (the number of infusion donor Lymphocyte was 4-8 ×108/kg) ,Cy, FLU, BU and TT started on day-8~-3. Unrelated cord blood was infused on day 0.GVHD prophylaxis consisted of FK506 and Mycophenolate Mofetil (MMF). Unrelated cord blood matching requires ≥7/10 HLA locus , and Kir assay was carried out at the same time. Cord blood with Haplotype Cen-Bx was preferred. Results: The median follow-up time was 32 months(2 to 52ms).47 patients were engrafted with cord blood in the two groups totally. In the CT group,Haplo-cells and UCB-cells engrafted finally in 24 and 31 patients, respectively.In the LCB group, only 1 of 17 cases developed primary graft failure,who achieved disease-free survival after the second haploid transplantation. Of the 47 patients with cord blood engraftment in the two groups, the median number of infusion TNC and CD34+ from cord blood were 8.1(3.3~17)×107/kg and 0.25(0.08~4.0)×106/kg, respectively. The median times to neutrophil more than 0.5×109/L and platelet more than 20×109/L were 38 (19-67) days and 36(12-67) days post-HSCT, respectively. The Overall survival (OS) and leukemia free survival (LFS) were 97.9% and 95.8%, respectively. There were no significant difference in the OS and LFS between CT group and LCB group (p=0.18 and p=0.50, respectively). The cumulative incidences of grades Ⅱ-Ⅳ° aGVHD was 29.8% (14pts).There were 4 patients with grade III° and Ⅳ° aGVHD ，all of whom were effectively improved. Chronic GVHD occurred in 13 patients who mainly had mild skin involvement. Only one of them had mild BO. The transplantation-related mortality was 2.1% (1 case died of severe pre-engraftment syndrome in LCB group). The transient reactivation of virus was mainly caused by HHV-6. One case developed HHV-6 virus encephalitis.And no VOD occurred. Summary: UnrelatedCord blood engraftment has significant clinical efficacy in the treatment of JMML,and it is relatively safe and reliable.UnrelatedCord blood could be used as a priority donor for such children. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Detection of Human Herpes Virus-6 in saliva of Patients with Bell's palsy

Background: Bell’s palsy is unilateral peripheral paralysis of the seventh cranial nerve, several mechanisms have been proposed in the pathogenesis of this disease, among the viral infections specially herpes virus's family including roseola viruses that have been detected in patients' saliva.&#x0D; Method: A case-control study conducted on Bell’s palsy patients at al-Imamein Al-kadhimein Medical City throughout the period from March 2019 to November 2019. Saliva samples were taken from 50 patients (18-55) years of age with early stages of Bell's palsy and from 50 apparently healthy and age and sex matched volunteers as control. Viral DNA was extracted from the saliva and then real time PCR for detection and quantification of HHV6 DNA in these patients&#x0D; Results: Saliva samples taken from 28 patients in the Bell’s palsy patients (56%) tested positive for HHV6, as vs. to just 8 (16%) in the healthy group(control) (P= 0.028). There was increase amount of HHV6 load in patients compare to control group (p=0.001). Regarding the demographic values, an important difference in the detection rate for Human herpes virus-6 between male (64%) and females (35%) (P= 0.02). There was significant correlation between age and grading (p =0.01), as increase age correlate with high grading, also between viral load of HH6 and grading as increase viral load of the virus correlate with high grading (p = 0.001).&#x0D; Conclusion: HHV-6 virus might have a potential role in the pathogenesis of Bell’s palsy.

Post-Transplant Cyclophosphamide, Abatacept, and Short Course of Tacrolimus Combination (CAST) Is Safe and Seems Highly Effective in Preventing Graft-Versus-Host Disease Following Haploidentical Peripheral Blood Stem Cell Transplantation

The introduction of post-transplant cyclophosphamide (PTCy) has circumvented the need for T-cell depletion following haploidentical stem cell transplantation (SCT). By expanding the donor pool for patients from certain ethnic minorities, this has addressed to some degree an important health care disparity issue in SCT. However, a recent registry study showed increased incidence GvHD and inferior outcomes in patients receiving haploidentical SCT with PTCy, tacrolimus and mycophenolate mofetil for GvHD prevention as opposed to matched unrelated donor SCT with PTCy-based GvHD prevention. Seeking to improve the results of GvHD prevention in the setting of haploidentical SCT, we examined a combination of PTCy, abatacept and a short course of tacrolimus (CAST). Abatacept is a recombinant soluble fusion protein composed of the extracellular domain of cytotoxic T-lymphocyte associated antigen-4 (CTLA-4) fused to the Fc region of IgG1. Abatacept blocks CD28-CD80I86 axis and prevents T-cell co-stimulation. In early studies, abatacept has shown promising results when added to methotrexate and tacrolimus in matched and mismatched donor SCT. We initiated a phase Ib-II clinical trial for patients with hematological malignancies undergoing haploidentical SCT. Patients received G-CSF mobilized peripheral blood grafts from related haploidentical donors. GvHD prevention consisted of PTCy 50mg/kg IV on day +3 and +4 with forced hydration, abatacept 10mg/kg IV on day +5, +14 and +28 and tacrolimus. Tacrolimus was started on day +5 at 0.02mg/kg/day by continuous IV and adjusted thereafter to maintain a trough level of 5-12ng/mL. Tacrolimus taper was planned to begin on day +60 and complete by day +90 in the absence of GvHD. All patients received standard supportive care including levofloxacin until neutrophil engraftment, posaconazole until day +75, acyclovir for 1 year and, if CMV positive by serology, letermovir until day +100. Pneumocystis Jiroveci prophylaxis was started after neutrophil engraftment and continued until 6 months post-transplant. G-CSF was administered routinely until neutrophil engraftment.Since September 2020, 19 patients were enrolled. Three patients are too early in their post-transplant course and were excluded from this analysis. Patients' characteristics are summarized in the table. All but 2 patients received cryopreserved products. Median times to ANC and platelet engraftment were 18.5 days (14-30) and 28.5 (16-61). All 16 patients achieved full whole blood donor chimerism by day +30. There was no secondary graft failure. With a median follow-up was 149.5 days (41-308) with 10 patients having >120 days and 8 >180 days of follow-up, 4 patients developed skin acute GvHD (all grade I). No patient developed grade II-IV acute GvHD. Two patients developed skin chronic GvHD (limited, both moderate). Both cases were diagnosed following COVID-19 vaccination. Fifteen patients completed tacrolimus taper by day +90. Two patients received systemic steroids, one for treatment of cGvHD. The remaining patients required no further immunosuppressive therapy beyond day +90. CMV activation rate was 25%. One patient had EBV reactivation and required preemptive therapy with 2 weekly rituximab doses. There were no cases of adenovirus, HHV-6 virus or BK virus reactivation. Four patients developed renal insufficiency (3 in the setting of acute sepsis and 1 with thrombotic microangiopathy, which resolved after tapering off tacrolimus. One patient with adult T-cell leukemia/lymphoma relapsed and died. All other patients are alive and well.In summary, our preliminary results suggest that CAST with shortened course of tacrolimus is feasible and seems to offer very promising outcomes with low rates of acute GvHD. The study is accruing actively and the results of a larger cohort with longer follow-up will be presented at the meeting. If confirmed, by improving the outcomes of haploidentical SCT, this regimen may further address a health care disparity issue, offering almost every patient in need of allogeneic SCT an alternative donor option with equal outcomes. [Display omitted] DisclosuresAl-Homsi: Daichii Sanyko: Consultancy; Celyad: Other: Advisory Board. Abdul-Hay: Abbvie: Consultancy; Servier: Other: Advisory Board, Speakers Bureau; Jazz: Other: Advisory Board, Speakers Bureau; Takeda: Speakers Bureau; Amgen: Membership on an entity's Board of Directors or advisory committees. OffLabel Disclosure:Abatacept - off label use as GvHD prevention Cyclophosphamide - off label use as GvHD prevention

SEASONAL FACTOR AND FREQUENCY OF DETECTION OF HERPES VIRUSES AND INFLUENZA VIRUS IN PATIENTS WITH ISCHEMIC STROKE

The aim: To determine the frequency of HSV1, HSV2, VZV, CMV, EBV, HHV6 and influenza virus detection in patients with ischemic stroke in different seasons. Materials and methods: 144 patients with ischemic stroke were examined: 78 (54.2%) women and 66 (45.8%) men, mean age of 63.1 ± 0.8 years. Detection of the herpesvirus DNA and the influenza virus RNA was performed using PCR monthly in 12 patients. Results: A manifestation of a viral infection was detected in 32 (22.2%) and virus genomes were observed in 29 (90.6%) patients. Viral infection frequency is significantly lower in summer, compared to winter-autumn; p=0.033. HSV1 and HHV6 were the most common (19 (52.8%) and 16 (44.4%)); VZV was the least common (5 (13.9%)). Influenza virus RNA was detected in 10 (27.8%) patients. In winter-autumn the frequency of HSV1, HSV2, HHV6 viruses detection is significantly higher, compared to the spring-summer (p<0.05), and the difference is almost significant for the influenza virus (p=0.060) and the EBV (p=0.060). Association of stroke occurrence with the presence of two or more types of viruses is more common in winter, compared to the summer season: 11 (30.6%) vs. 3 (8.3%), p=0.017. Conclusions: Prevention and treatment of herpesvirus infections exacerbations, in particular HSV1 and HSV2, which significantly increase in winter, compared to summer, is an important direction of stroke prevention measures in risk groups.

Viral serology in “infarct-like” acute myocarditis: Useful or useless?

The objectives of this study were to assess the viral serology of the patients with acute myocarditis and the relation with clinical and CMR parameters at the acute phase and at follow-up. All cases of infarct-like acute myocarditis, revealed as an acute coronary syndrome with chest pain, elevated troponin levels and confirmed by cardiac magnetic resonance, from 2012 to 2018, were included at Dijon University Hospital. A blood sampling including viral serology is obtained at the time of diagnostic. One-year follow-up included a complete clinical and rhythmic evaluation, and a CMR control. Among the patients with “infarct like” acute myocarditis, serological analyses were performed in 62 patients. Regarding cardiotropic viruses particularly detected in myocarditis, HHV6 is found in 25 patients (40%), Parvovirus B19 is found in 2 patients (6%) and Coxsackie virus is found in 2 patients (4%). No statistical difference was observed regarding demographic and clinical parameters according the presence or absence of cardiotropic viruses. However, concerning CMR parameters, the extent of myocarditis is more important in the “cardiotropic viruses” group at the acute phase ( P = 0.05) and at three months ( P = 0.007) compared to the “no cardiotropic viruses” group (CV+ group: SQS Score in acute phase = 10,4–SQS Score at 3 months = 6,3 VS CV- group: SQS Score in acute phase = 7,3–SQS Score at 3 months = 2,7). At one year, there was no significant difference between the two groups regarding CMR parameters. Cardiotropic viruses (HHV6, Parvovirus B19 and Coxsackie virus) were found in almost 50% of cases in “infarct like” acute myocarditis. According the presence or absence of cardiotropic viruses, CMR parameters were different. Furthermore, use of quantitative PCR (detection of viral genomes) or detection of viral DNA and RNA via in situ hybridization would be a challenge for the future identification of viruses in myocarditis.

Genome-wide analysis of codon usage pattern in herpesviruses and its relation to evolution

The preferential use of a specific codon, out of a group of synonymous codons encoding the same amino acid, in a gene transcript results from the bias in codon choice. Various evolutionary forces namely mutation pressure and natural selection influence the pattern of codon usage i.e. distinct for each gene/genome. We investigated the pattern of codon usage of eight human herpesvirus genomes and compared them with two other herpesvirus genomes namely murine herpesvirus 68 and bovine herpesvirus type 1.1 to elucidate its compositional features, pattern of codon usage across the genomes and report the differences of codon usage pattern of human herpesviruses from that of other two other viruses. We also identified the similarity of the codon usage of human herpesviruses with its host (human). The genes were found to be CG rich in HHV2, HHV3, HHV4, HHV6, HHV7 and BH genomes while TA rich in HHV1, HHV5, HHV8 and MH genomes. The codon usage bias (CUB) of genes was low. A highly significant correlation was found among compositional contents depicting the role of mutational pressure along with natural selection in framing CUB. Several more frequently used codons as well as less frequently used codons were identified to be similar between each human virus and its host (human), while murine herpesvirus 68 and bovine herpesvirus type 1.1 genomes did not possess similar adaptation strategy as human herpesviruses to human (host), thus we could conclude that viral CUB might have been shaped as per their host’s nature for better surveillance. Neutrality plot revealed mutational pressure mostly influenced the CUB of HHV1, HHV8 and MH viruses, while natural selection had a major impact in the CUB of HHV2, HHV3, HHV4, HHV5, HHV6, HHV7 and BH genomes.

HIV-Related Lymphoproliferative Diseases in the Era of Combination Antiretroviral Therapy.

HIV-positive patients have a 60- to 200-fold increased incidence of Non-Hodgkin Lymphomas (NHL) because of their impaired cellular immunity. Some NHL are considered Acquired Immunodeficiency Syndrome (AIDS) defining conditions. Diffuse large B-cell Lymphoma (DLBC) and Burkitt Lymphoma (BL) are the most commonly observed, whereas Primary Effusion Lymphoma (PEL), Central Nervous System Lymphomas (PCNSL), Plasmablastic Lymphoma (PBL) and classic Hodgkin Lymphoma (HL) are far less frequent. Multicentric Castleman disease (MCD) is an aggressive lymphoproliferative disorder highly prevalent in HIV-positive patients and strongly associated with HHV-8 virus infection. In the pre-Combination Antiretroviral Therapy (CART) era, patients with HIV-associated lymphoma had poor outcomes with median survival of 5 to 6 months. By improving the immunological status, CART extended the therapeutic options for HIV positive patients with lymphomas, allowing them to tolerate standard chemotherapies regimen with similar outcomes to those of the general population. The combination of CART and chemotherapy/ immuno-chemotherapy treatment has resulted in a remarkable prolongation of survival among HIV-infected patients with lymphomas. In this short communication, we briefly review the problems linked with the treatment of lymphoproliferative diseases in HIV patients. Combination Antiretroviral Therapy (CART) not only reduces HIV replication and restores the immunological status improving immune function of the HIV-related lymphomas patients but allows patients to deal with standard doses of chemotherapies. The association of CART and chemotherapy allowed to obtain better results in terms of overall survival and complete responses. In the setting of HIV-associated lymphomas, many issues remain open and their treatment is complicated by the patient’s immunocompromised status and the need to treat HIV concurrently.

Клінічні та УЗДГ кореляції у хворих на ішемічний інсульт iз маніфестацією вірусної інфекції в анамнезі

The goal. The goal of the work was to study the structure of viral and non-viral infections in patients with ischemic stroke, as well as the relationship between neurological recovery and severity of cerebral atherosclerosis, depending on the type of infectious agent.Materials and methods. A total of 89 patients with ischemic acute cerebral circulation disorder and a history of clinical manifestations of a viral infection in the 3 months prior to the cerebrovascular accident were included in the study. Among the patients 44 (44.9%) were men and 54 (55.1%) were women with an average age of 62.08 ± 1.29 years (from 36 years to 92 years). 61 (81.9%) patients were diagnosed with ischemic stroke, and 28 (18.1%) patients were diagnosed with TIA.The diagnosis was confirmed by magnetic resonance imaging (MRI), and the severity of neurological status was assessed using the National Institute of Health Stroke Scale (NIHSS) on the first, 7th and 21st day. Patients' blood serum was examined for the presence of herpes viruses DNA, the RNA of influenza viruses, enteroviruses, the viruses of an acute respiratory infection (adenoviruses), as well as for the common non-viral infections: mycoplasma, ureoplasma, toxoplasma, and chlamydia. The method of polycytomerase chain reaction (PCR) for DNA and RNA viruses extraction from blood serum (6; 8) and the method of enterovirus antigens detection in a cell culture by enzyme-linked immunosorbent assay (ELISA) (21; 24; 40) were used.The degree of cerebral atherosclerosis was determined by ultrasound duplex scanning (UDS) of brachiocephalic arteries.Methods of descriptive statistics were used.Research results. The average period from the clinical manifestation of the infection to the development of stroke in patients with viral persistence was 11.8 ± 2.5 days (from 1 day to 90 days). Up to 7 days — 32 (36.0%) patients, from 7 days to 14 days inclusive — 44 (49.4%), more than 14 days — 13 (14.6%).There was a weak but definite inverse correlation between the term of the viral manifestation and the severity of the acute cerebrovascular accident (r = 0.237, p = 0.025).HSV1 (51) was detected most often in the acute period of stroke in 57% of patients, (p <0.05). The persistence of HSV2 and HHV6 was 38.2% and 32.6% respectively. A combination of two different HSV2 and HHV6 viruses was observed in 84 (89.9%) patients.The combination of HSV1 / HSV2 is most commonly observed (in 29 (32.6%) patients p <0.05). Coefficients of correlation between the presence of certain types of viruses, the number of stenoses, and the number of hemodynamically significant stenoses ranged from 0.19 with ARI to 0.33 in the case of a combination with HHV6.There was no detectable relationship between the severity of neurological deficiency on the first day after the stroke and the type of viral infection or a combination thereof, except for adV (r = 0.27, p = 0.01).The reduction in the NIHSS score on the 7th and the 21st day negatively correlated with the presence of certain viruses and their combinations, in particular herpesviruses or their associations HSV1, HSV1,2; HSV1,2-HHV6.Conclusion. The DNA of HSV1, HSV2, HHV6, HHV4, HHV5 viruses was most often found in patients with ischemic stroke and a clinical manifestation of a persistent viral infection in history (within 1-90 days before the development of symptoms),The presence of major vessels stenosis in the extracranial unit, the number of stenoses, and the presence of hemodynamically significant stenoses correlated with the presence of viral persistence of HHV6 and / or the association of HHV6 and influenza.The presence of HSV1, HSV2, HHV6 viruses and / or their associations worsens the restoration of neurological functions in the acute post-ischemic period setting.

Virus persistence in atherosclerotic plaques in ischemic stroke/TIA patients with carotid endarterectomy

Objective — to assess the frequency of detection of embologic and nonembologenous atherosclerotic plaques, moderate stenosis (50 — 69 %) and severe degree (79 — 99 %) in patients with ischemic stroke and transient ischemic attack with progression of atherosclerosis and the presence of herpetic infection of the carotid endothelium.Methods and subjects.103 patients with IS/TIA after carotid endothelium were examined: 67 males of 66.1 ± 1.4 (42 — 82 y/o), 36 females 63.0 ± 1.3 (44 — 81 y/o). Mean age was 65.1 ± 0.9. We examined the width of IMC, presence of the atherosclerotic plaques, their constitution (hypoechoic, echoic), structure, localization, spread, shape. Using the methods of transfection and polymerase chain reaction in the atherosclerotic plaque and blood of patients, the presence of HSV1, 2, EBV, CMV, HHV6 viruses was established.Results. In 32 patients in atherosclerotic plaque and blood, prevalence of CMV and its viral atherosclerosis associations (CMV/HSV1, CMV/HSV2) was revealed, in 23 — HHV6 isolated and in atherosclerosis associations (HSV1/HHV6, HHV6/HSV1/HSV2).Conclusions. In the case of persistence of CMV and atherosclerosis, CMV/HSV1 and CMV/HSV2 significantly increase the risk of developing hypoechoic embolic atherosclerotic plaques in the internal carotid artery. In the case of persistence of HHV6, the association of HSV1/HHV6 and HSV1/2/HHV6 in atherosclerotic plaques in the internal carotid arteries, the relative risk of developing severe stenosis in patients with symptomatic stenosis is significantly increased, which is an indication for carotid endothelium. The results obtained testify to the need for periodic ultrasound scan to exclude stenosis of the internal carotid artery in patients with cerebrovascular disease.

Allogeneic stem cell transplant (HSCT) for acute lymphoblastic leukaemia (ALL) using CD34 selected stem cells followed by prophylactic infusions of pathogen-specific and CD19 CAR T cells

Background & Aim Disease relapse, graft versus host disease (GVHD) and infection are responsible for the majority of deaths after HSCT. We assessed a cellular engineering approach to simultaneously reduce the incidence of these complications using CD34+ stem cell selection combined with post-transplant infusion of pathogen-specific and leukemia-specific T cells. Methods, Results & Conclusion METHODS We conducted a pilot study in which patients with ALL in remission were treated with a CD34-selected stem cell transplant followed by two planned prophylactic infusions of donor-derived T cells targeting opportunistic pathogens (CMV, EBV and Aspergillus), and leukaemia (CD19 CAR T cells). Cellular products were manufactured locally. Patients did not receive anti-thymocyte globulin or post-transplant GVHD prophylaxis. RESULTS We performed matched sibling donor transplants on two patients aged 45 and 27 years with ALL in morphological CR. Patient 1 had Ph1 positive ALL with detectable BCR-ABL at the time of transplant (0.001%). Patient 2 was Ph1 negative and had no detectable disease by flow cytometry pre-transplant. Conditioning was cyclophosphamide 120mg/kg and TBI 1200cGy. Patients received CD34 selected stem cells (total CD34+ doses 3.5 and 3.6 × 106/kg; total CD3+ cell doses 1.3 and 0.2 × 104/kg). Neutrophil engraftment (>0.5) occurred on days 11 and 12, platelet engraftment (>20) on days 8 and 13. Patients received infusions of pathogen specific T cells (day 21) and CD19 CAR T cells (days 27 and 21). CRS (grades 1 and 2) and neurotoxicity (grade 1) developed in both patients; treated with tocilizumab, patient 1 also received dexamethasone. CAR T cells proliferated in vivo despite low disease burden and persisted in blood for at least 6 weeks in both patients (Figure 1). Patient 1 received foscarnet for 9 days and a second pathogen-specific T cell infusion on day 47 post-transplant. Patient 2 developed asymptomatic reactivation of HHV6 and BK virus. At 364 and 350 days post-transplant, both patients are well, free of GVHD and remain in remission. CONCLUSION This is the first description of a strategy that combines prophylactic donor-derived CAR T cell and pathogen-specific T cell administration after HSCT in the context of CD34+ selection to minimise development of GVHD. Our results demonstrate absence of GVHD, minimal treatment for viral reactivation and remission at 12 months post-transplant suggesting that this approach has promise for improving GVHD relapse free survival following HSCT.

Transplantation Using Bone Marrow from a (very) HLA Mismatched Unrelated Donor in the Setting of Post-Transplant Cyclophosphamide Is Feasible and Expands Access to Underserved Minorities

Background Despite increasing donor options for allogeneic transplantation, including matched/mismatched related donors, matched unrelated donors and cord blood units, a proportion of patients do not find a donor. This is especially relevant in patients from racial/ethnic minorities. Post-transplant cyclophosphamide (PTCY) has successfully overcome barriers related to HLA-mismatching in the related donor setting. We hypothesized that transplantation with a mismatched unrelated donor (MMUD) using PTCY would be feasible and associated with high engraftment and acceptable GVHD incidence. Methods We performed a prospective Phase II study of MMUD bone marrow (BM) transplantation with PTCY for patients with hematologic malignancies. Patients with a suitable HLA matched related or URD were excluded. Patients received a fresh BM graft, followed by PTCY on days +3, +4, Sirolimus/MMF starting on Day+5. Matching for 4-7/8 at HLA-A, -B, -C, and –DRB1 was permitted. We enrolled 80 patients (40 full intensity conditioning [FIC]; 40 reduced intensity conditioning [RIC]) at 11 transplant centers in the U.S. between Dec 2016 and March 2019. Regimen intensity was at the center's discretion. Results Characteristics are shown in Table 1. Importantly, 48% of patients were non-white/Hispanic, 55% had an HCT-CI >2 and 34% had a KPS of 1 HLA allele, 59% were under 30. Overall survival and non-relapse mortality at 100 days were 92% and 5% in the FIC arm, and 90% and 7.5% in the RIC arm (Table 2). Neutrophil recovery was 98% in both arms, with no primary graft failure in the FIC arm, and 7.5% in the RIC arm. Peripheral blood donor chimerism was >75% at all timepoints. Acute GVHD grade III-IV at day 100 was relatively high at 25% in the FIC arm but very low at 2.6% in the RIC arm. Viral reactivations were high (Table 3). Conclusion These early (day 100) results of our prospective study show high rates of engraftment with acceptable rates of GVHD in recipients of MMUD transplantation with PTCY, despite a high degree of HLA mismatch. An important finding is that ethnic minorities represent almost 50% of the study population, showing that this approach expands access to patients in need of potentially curative therapy. BK and HHV-6 virus reactivation/infection were common, the clinical significance of which requires further study. Understanding the risks for and impact of graft failure and acute GVHD is being studied as follow up continues.

Delayed CD4+ T-Cell but Faster B-Cell Immune Reconstitution after Ptcy-Based Compared to Conventional Gvhd Prophylaxis after Allogeneic Transplantation

<h3>Background</h3> Post-transplant cyclophosphamide (PTCY) is being increasingly used for graft-versus-host disease (GVHD) prophylaxis after hematopoietic cell transplantation (HCT) across various donor types. However, immune reconstitution after PTCY-based vs. conventional GVHD prophylaxis has not been well studied. <h3>Methods</h3> We evaluated the pace of immune reconstitution (CD4+ T-cell, CD8+ T-cell, NK-cell and B-cell) at 3 months, 6 months and 1 year post-HCT in 583 adult patients receiving myeloablative (n=223) or reduced intensity conditioning (n=360) HCT (2012-2018). Haploidentical (Haplo; n=75) and 8/8 HLA-matched unrelated (MUD, n=508) donor types were included. GVHD prophylaxis was PTCY-based in 75 Haplo and 38 MUD (MUD-PTCY) HCT, while tacrolimus/methotrexate (TAC/MTX) was used in 89 and TAC/Sirolimus (TAC/SIR) in 381 MUD HCT. Clinical outcomes including viral infections, non-relapse mortality (NRM) and overall survival (OS) were compared across all four treatment groups. <h3>Results</h3> The recovery of absolute total CD4+ T-cell count after Haplo-PTCY and MUD-PTCY was significantly lower compared to MUD TAC/MTX or MUD TAC/SIR throughout 1 year of HCT (Figure). In contrast, CD19+ B-cell recovery at 6 months and thereafter was more rapid after Haplo-PTCY and MUD-PTCY HCT in comparison to MUD TAC/MTX and MUD TAC/SIR. In subgroup analysis compared to MUD TAC/MTX HCT, total CD8+ T-cell or NK-cell recovery was not significantly different after Haplo-PTCY or MUD-PTCY HCT. However, patients receiving MUD TAC/SIR vs. MUD TAC/MTX had significantly slower reconstitution of total CD8+ T-cells up to 6 months and CD19+ B-cells at 1 year of HCT, but no significant differences in CD4+ T-cell or NK-cell recovery. The distribution of recipient CMV serostatus was similar in all four groups: 72% in Haplo-PTCY, 62 % in MUD-PTCY HCT, 68% in MUD TAC/MTX and 63% in MUD TAC/SIR (<i>p</i>=0.11). Cumulative incidence of CMV reactivation/infection at 1-year of HCT was higher in patients receiving Haplo-PTCY (39.6%) or MUD TAC-MTX (37.7%) compared to those receiving MUD-PTCY (27.0%) or MUD TAC/SIR (22.8%; <i>p</i><0.01). Whereas the incidence of EBV reactivation/infection was lower in patients receiving PTCY-based GVHD prophylaxis (5.2% in Haplo-PTCY and 8.1% in MUD-PTCY) as compared to patients receiving MUD TAC/MTX (19.7%) or MUD TAC/SIR (20.5%, <i>p</i><0.01). The incidences of HHV6, BK- and other viruses were similar between the groups. Overall, the treatment groups had comparable NRM (<i>p</i>=0.27) and OS outcomes (<i>p</i>=0.78). <h3>Conclusion</h3> Our data demonstrate that the pattern of immune reconstitution after HCT is different after PTCY-based vs. conventional GVHD prophylaxis with delayed total CD4+ T-cell but more rapid B-cell recovery after PTCY-based GVHD prophylaxis. The rates of CMV and EBV viral infections were different across the donor types. However, these differences had no significant influence on NRM or survival after HCT.

Viral serology in “infarct-like” acute myocarditis: Useful or useless?

The most common cause of myocarditis is viral infection, especially from cardiotropic viruses (Parvovirus B19, Human Herpesvirus 6 and Coxsackie) resulting in a destruction of the myocardium. Moreover, a serological analysis is not systematic and their value remains unclear. The objectives of this study were to assess the viral serology of the patients with acute myocarditis and the relation with clinical and CMR parameters at the acute phase and at distance. All cases of infarct-like acute myocarditis confirmed by CMR from April 2012 to January 2018 were included in a prospective study at Dijon University Hospital. A blood sampling including viral serology is obtained at the moment of the diagnostic. One-year follow-up included a complete clinical and rhythmic evaluation, and a CMR control. Among the patients with “infarct like” acute myocarditis, serological analyses were performed in 62 patients and cardiotropic viruses are found in 29 patients. No statistical difference was observed regarding demographic and clinical parameters. However, concerning CMR parameters, the extent of myocarditis is more important in the “cardiotropic viruses” group at the acute phase (10.4 vs. 7.3 – P = 0.05) and at three months (6.3 vs. 2.7 – P = 0.007) compared to the “no cardiotropic viruses” group. At one year, there was no significant difference between the two groups regarding CMR parameters. Cardiotropic viruses (HHV6, Parvovirus B19 and Coxsackie virus) were found in almost 50% of cases in “infarct like” acute myocarditis. According the presence or absence of cardiotropic viruses, CMR parameters were different. Furthermore, use of quantitative PCR (detection of viral genomes) or detection of viral DNA and RNA via in situ hybridization would be a challenge for the future identification of viruses in myocarditis.

Mathematical Model of Classical Kaposi’s Sarcoma

In this paper, the global properties of a classical Kaposi’s sarcoma model are investigated. Lyapunov functions are constructed to establish the global asymptotic stability of the virus free and virus (or infection) present steady states. The model considers the interaction of B and progenitor cells in the presence of HHV-8 virus. And how this interaction ultimately culminates in the development of this cancer. We have proved that if the basic reproduction number, R0 is less than unity, the virus free equilibrium point, e0, is globally asymptotically stable (GAS). We further show that if R0 is greater than unity, then both the immune absent and infection persistent steady states are GAS.

A Meta-Analysis of Genetic Abnormalities and Next Generation Sequencing of Primary Cases of Castleman Disease

Castleman disease is a rare lymphoproliferative disorder known to represent at least four distinct clinico-pathologic sub-types. Large advancements in our clinical and histopathologic description of these diverse diseases have been made, resulting in subtyping based on degree of nodal involvement (unicentric versus multicentric) and according to viral infection (human herpes virus 8 - HHV8 and Human Immunodeficiency Virus - HIV). However, we continue to lack a foundational understanding of the biologic mechanisms driving at least a subset of patients with these diseases. In recent years, significant molecular and genetic abnormalities that may be associated with this disease have been described; yet we are still unclear as to a unified vision of what loci and genes are involved in at least a subset of cases of Castleman disease. In our study we performed a meta-analysis of all cases (unicentric and multicentric) of Castleman disease described in literature to date, and correlate cytogenetic, molecular and genetic abnormalities with disease subtypes, phenotypes and clinical outcomes. We further performed targeted DNA deep sequencing and RNA-sequencing on a cohort of Castleman Disease cases. Our results demonstrate that genetic abnormalities in the MAPK-pathways, PI3K-pathway and Interleukin signaling are frequent in unicentric Castleman disease, whereas abnormalities in DNA methylation are seen more commonly in idiopathic multicentric Castleman disease, along with abnormalities in MAPK-pathways. We conclude that UCD and iMCD may share similar pathways of MAPK pathogenesis but can also be distinguished by cytokine and other DNA modification pathways that differ. Disclosures Ohgami: Agilent technologies: Other: received support/funding.