anti-CMV Antibodies Research Articles

Seroprevalence and Molecular Detection of Cytomegalovirus UL146 and US28 Gene Expression in Women With Recurrent Pregnancy Loss.

Background Human cytomegalovirus (CMV) is a global herpesvirus that is highly prevalent worldwideand is able to establish lifelong latency after initial infection. The infection is highly frequent during pregnancy in human beings, which leads to preterm birth in some cases. Circulating strains of CMV carry a high number of variable or disrupted genes. Some of these like UL146, a highly diverse gene, and theUS28 gene are involved in viral dissemination. This study aims to determine the seroprevalence of CMVand to investigate whether the highly variable UL146 and US28 genes, isolated from the blood of seropositive women, are associated with recurrent pregnancy loss. Material and methods This cross-sectional study was carried out in Erbil City, Iraq from October 2022 to July 2023.A total of 150 women at their reproductive age with a history of miscarriage who attended Maternity Teaching Hospital were enrolled. Anti-CMV IgG and IgM antibodies were assessed by enzyme-linked immunosorbent assay (ELISA). Highly variable UL146 and US28 genes of CMVfrom seropositive samples were amplified by conventional polymerase chain reaction (PCR), and the results were visualized on a UV-transilluminator. SPSS version 22(IBM Corp., Armonk, NY, USA)was used for data entry and analysis. The p-value less than 0.05 was regarded as statistically significant. Results Anti-CMV IgG and IgM were seropositive in 103 (53.3%) and 13 (8.7%) women, respectively, and only 10 (6.7%) of them for both anti-CMV IgG and IgM. Significant associations of CMV and history of miscarriage, age, educational level, and gestational age of miscarriages were observed(p-value less than 0.05). On the other hand, no statistically significant association between CMV and socioeconomic level or residency was observed. The frequencies of genetic analysis of UL146and US28 of the 103 seropositive tested samples of women with a history of miscarriage were 31 (30.1%)and nine (8.7%), respectively. A significant association between recurrent miscarriage and UL146 gene expression was observed. PCR targeting theUL146 demonstrated greater sensitivity for diagnosing CMV. Conclusion The seroprevalence of CMV is relatively high in Erbil, and the UL146 and US28 genes can act as factors in the initial level of CMV. Therefore, molecular detection of these genes can aid in determining the virulence of CMV strains.

Decreased T-cell response against latentcytomegalovirus infection does not correlate with anti-IFN autoantibodies in patients with APECED.

Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) is an inborn error of immunity affecting both multiple endocrine organs and susceptibility to candidiasis, each with an autoimmune basis. Recently, high titer neutralizing anti-type I interferon (IFN) autoantibodies have been linked with increased severity of SARS-CoV-2 and varicella zoster virus infections in APECED patients. Examining immunity against cytomegalovirus (CMV), we found a higher prevalence of anti-CMV IgG antibodies in patients with APECED (N = 19) than in 44 healthy controls (90% vs 64%, p = 0.04); the similar difference in their IgG levels did not achieve significance (95 ± 74 vs 64 ± 35 IU/mL, ns.). In contrast, the frequency of CMV-specific Tcells was lower (804 ± 718/million vs 1591 ± 972/million PBMC p = 0.03). We saw no correlations between levels of anti-CMV IgG and anti-IFN antibodies in APECED patients or in a separate cohort of patients with thymoma (n = 70), over 60% of whom also had anti-IFN antibodies. Our results suggest a dysregulated response to CMV in APECED patients and highlight immunodeficiency to viral infections as part of the disease spectrum.

Seroprevalence of ToRCH Pathogens among Children Admitted to a Tertiary Care Hospital in Eastern India for Cataract Surgery and Cochlear Transplantation.

The study aims to investigate the presence of TORCH infections in a child with bilateral cataracts and deafness and report the ToRCH-serology screening profile (Toxoplasma gondii (TOX), rubella (RV), cytomegalovirus (CMV), and herpes simplex virus (HSV-I/II)) in pediatric cataract and deafness. Cases that had a clear clinical history of congenital cataracts and congenital deafness were included in the study. The study population consisted of 18 bilateral cataracts and 12 bilateral deafness child who was admitted to AIIMS Bhubaneswar for cataract surgery and cochlear implantation, respectively. Sera of all children were tested qualitatively and quantitatively for IgG/IgM-antibodies against ToRCH agents in a sequential manner. Anti-IgG antibodies against the torch panel were detected in all cataract and deafness patients. Anti-CMV IgG was detected in 17 of 18 bilateral cataract children and 11 of 12 bilateral deaf children. The rates of anti-CMV IgG antibody positivity were significantly higher. In the cataract group, 94.44% and in the deafness group, 91.66% of the patient was Anti-CMV IgG positive. Besides this, 77.7 % of the patient from the cataract group and 75% from the deafness group was anti- RV IgG antibody positive. In bilateral cataract patients, IgG-alone seropositive cases were mostly attributed to CMV (94.44%; 17/18), followed by RV (77.70%; 14/18), HSV-I (27.70%; 5/18), TOX (27.70%; 5/18), and HSV-II (16.60%; 3/18). In bilateral deafness patients, the spectrum of IgG alone seropositive cases was almost the same except for TOX (0/12). The current study recommends interpreting ToRCH-screening in pediatric cataracts and deafness with caution. Interpretation should include both serial qualitative and quantitative assays in tandem with clinical correlation to minimize diagnostic errors. The sero-clinical-positivity needs to be tested in older children who might pose a threat to the spread of infection.

Epidemiological Insights into TORCH Infections in the Population of Erbil City

A seroepidemiological study of the TORCH panel was conducted on inhabitants of Erbil City to provide updated baseline data on TORCH prevalence. 508 individuals were included in this study. 218 (42.91%) were females and 290 (57.09%) were males. Their ages ranged from 13 to 63 year, the majority being within the age group (21-30 years), 162 (31.89%), (P < 0.05). 20 individuals (3.94%) tested positive for anti-TOX IgG antibodies. One of the 20 (5%) tested positive for anti-TOX IgM antibodies, who was a 32-year-old female. 128 individuals (25.2%) (all females) tested positive for anti-Rubella IgG antibodies. 40 individuals (7.87%) were positive for anti-CMV IgG antibodies. One of the 40 (2.5%) also tested positive for anti-CMV IgM antibodies, who was a 26-year-old male. 18 individuals (3.54%) tested positive for anti-HSV-1 IgG antibodies. One of the 18 (5.56%) also tested positive for anti-HSV-1 IgM antibodies. One 18-year-old male (0.2%) tested positive for anti-HSV-2 IgG antibodies, and none were positive for anti-HSV-2 IgM antibodies. The results suggest the presence of Toxoplasma, CMV, HSV-1 and HSV-2 infections among the community, the majority (P < 0.05) being CMV followed by TOX and HSV-1 and finally HSV-2. The positive IgG results of Rubella are most probably due to the obligatory vaccination program for females. Despite the fact that the majority of positive cases were for IgG, enhancing vaccination efforts and providing comprehensive health education is crucial for enhancing the well-being of the Erbil population.

Major low-energy trauma results in non-specific immunoglobulin generation without evidence for specific autoantibody production: A prospective cohort study.

Cellular debris resulting from large trauma might overwhelm the scavenger mechanisms and lead to autoimmune reactions. We analysed whether a major well-defined trauma in humans induces laboratory signs of transient autoimmunity in the months after the insult. We included 50 patients with pertrochanteric femur fracture undergoing intramedullary nail osteosynthesis in a prospective cohort study and followed them at 3-4 days, 6 weeks, 12 weeks and 12 months postoperatively. By standard techniques, we assessed levels of total immunoglobulins, anti-nuclear antibodies (ANA), anti-cardiolipin antibodies, anti-dsDNA antibodies and anti-C1q antibodies, as well as antibodies against cytomegalovirus (CMV) as a control. Blood leukocyte differential and lymphocyte subpopulations were determined at baseline and in the first two postoperative samples. The mean age of the patients reached 80.1 years, and 23 (46%) completed all visits. Serum concentrations of total IgG, IgM and IgA increased at all follow-up time points. The ANA fluorescence light intensity units increased at 12 weeks and 12 months postoperatively (p < 0.0001), but the proportion of ANA-positive patients did not change (35%). The values of anti-C1q mildly increased at all follow-up visits, but not the ratio to total IgG. Anti-dsDNA remained negative in all patients, and anti-cardiolipin IgG/IgM antibodies did not change. Anti-CMV IgG antibodies increased significantly at all follow-up visits, without change in the ratio to total IgG. Flow cytometry showed an increased proportion of B-cells 3-4 days postoperatively. In conclusion, major musculoskeletal trauma in elderly patients induces a generalized non-specific increase in immunoglobulin production without laboratory signs for enhanced systemic autoimmunity.

Anti-cytomegalovirus antibody levels stratify human immune profiles across the lifespan.

Human cytomegalovirus (hCMV) is a ubiquitous latent persistent herpesvirus infecting 60-90% of the population worldwide. hCMV carriage in immunocompetent people is asymptomatic; thus, hCMV can be considered a component of normative aging. However, hCMV powerfully modulates many features of the immune, and likely other, systems and organs. Questions remain as to how hCMV carriage affects the human host. We used anti-CMV antibody titers as a stratifying criterion to examine the impact of "intensity" of hCMV infection as a potential biomarker of aging, inflammation, and immune homeostasis in a cohort of 247 participants stratified into younger (21-40years) and older (> 65years of age) groups. We showed that anti-CMV antibody titers increased with age and directly correlated to increased levels of soluble tumor necrosis factor (sTNFR) I in younger but not older participants. CD8 + cell numbers were reduced in the older group due to the loss in CD8 + T naïve (Tn) cells. In CMV carriers and, in particular, in anti-CMV Ab-high participants, this loss was mitigated or reversed by an increase in the numbers of CD8 + T effector memory (Tem) and T effector memory reexpressing CD45RA (Temra) cells. Analysis of CD38, HLA-DR, and CD57 expression revealed subset (CD4 or CD8)-specific changes that correlated with anti-CMV Ab levels. In addition, anti-CMV Ab levels predicted anti-CMV CD8 T cell responsiveness to different CMV open reading frames (ORFs) selectively in older participants, which correlated to the transcriptional order of expression of specific CMV ORFs. Implications of these results for the potential predictive value of anti-CMV Ab titers during aging are discussed.

Seroprevalence of Anti-Cytomegalovirus Antibodies in Pregnant Women from South-West Romania.

Cytomegalovirus (CMV), in addition to other agents, is part of the TORCH complex (Toxoplasma gondii, Rubella virus, Cytomegalovirus, Herpes simplex viruses, and other agents). CMV infection is the most frequent cause of congenital malformations. This study aimed to establish the variation of prevalence of anti-CMV antibodies in pregnant women from the South-West region of Romania, according to demographic factors, such as age and area of residence, in two separate time periods (2013-2016 and 2019-2022). We collected from the hospital records the age, place of residence, and anti-CMV antibody test results using immune electrochemiluminescence and chemiluminescence. This study found that the seroprevalence of anti-CMV IgM antibodies increased slightly from 2013-2016 to 2019-2022, from 1.92% to 2.26%, and for IgG antibodies from 93.68% to 94.96%. In both groups was observed a descending trend of anti-CMV IgM seroprevalence with an increase in age, showing a decrease in seroprevalence from 3.57% to 1.09% in pregnant women from rural areas in the 31-35 years age group, while in urban areas, we observed a decrease in seroprevalence from 11.11% to 3.06% in the <20 years age group. The IgG seroprevalence showed an increase both in rural areas (from 93.97% to 95.52%) and urban areas (from 93.52% to 94.27%). In both groups, seroprevalence was higher in rural areas compared to urban regions. These results show a high rate of immunization against CMV in pregnant women in South-West Romania, which led to a low risk of acquiring the primary infection during pregnancy. However, the increase in the rate of primary CMV infections in pregnancy suggests the need for prioritizing screening programs and improving the existing protocols to enhance maternal and child healthcare.

Frequency of Toxoplasma gondii, Rubella, and Cytomegalovirus Antibodies in Pregnant Women in Yazd Province, Iran

Background: TRC (Toxoplasma gondii, rubella virus, and cytomegalovirus [CMV]) infections during pregnancy can lead to serious sequelae in the uterine fetus. Prenatal testing, which includes screening and diagnosis of antibodies to TRC infections, is one of the most important aspects of prenatal care recommended during pregnancy. The aim of the present study was to determine the seroprevalence of TRC infections in pregnant women who attended the antenatal clinic in Yazd province in central Iran. Methods: For this purpose, 8355 sera samples were obtained from pregnant women attending the central laboratory of Yazd province in central Iran and subjected to screening for immunoglobulin G (IgG) and IgM antibodies against TRC using the enzyme-linked immunosorbent assay method. Of these, 4,245, 2,190, and 1,920 pregnant women were screened for anti-Toxoplasma, rubella, and CMV, IgG, and IgM antibodies, respectively. Results: The results revealed that 77.4% (1695/2190) and 78.1% (1500/1920) were positive for anti-rubella and anti-CMV IgG antibodies, and n.o seropositive was detected for anti-rubella and -CMV IgM antibodies. For anti-Toxoplasma antibodies, 20.85% (885/4245), 9.54% (405/4245), and 12.01% (510/4245) were positive for IgG, IgM, and both IgG-IgM antibodies, respectively. In addition, our findings showed the high prevalence of chronic TRC infections and a low recently acquired Toxoplasma infection in pregnant women. Conclusion: Our findings confirmed the high prevalence of chronic TRC infections and a low recently acquired Toxoplasma infection in pregnant women. Hence, observing personal behavioral practices (the source of infection and hygienic measures) are recommended to women at reproductive-age in general and seronegative pregnant women in particular.

Low-Dose ATG Has the Same Risk of CMV Reactivation after Allogeneic PBSCT As Ptcy, but Letemovir Is Effective in Preventing CMV Reactivation

[Introduction] Allogeneic peripheral blood stem cell transplantation (allo-PBSCT) is a curative treatment for hematological malignancies but has a risk of a chronic graft-versus-host disease (GVHD) compared to other donor sources. Post-transplant cyclophosphamide (PTCY) has been spread rapidly worldwide from HLA haploidentical-SCT (Haplo-SCT) to HLA matched-SCT. Although PTCY was theoretically thought to preserve non-alloreactive T cells which can contribute to fight infection, recent studies showed that PTCY was associated with the increased incidence of cytomegalovirus (CMV) infection. Letermovir (LTV), a novel anti-CMV agent, was useful for the prevention of CMV reactivation after SCT including Haplo-SCT using PTCY. On the other hand, low-dose antithymocyte globulin (ATG) is an alternative and useful option for GVHD prophylaxis in allo-PBSCT and often used in Japan. However, the risk of post-transplant CMV reactivation and the protective effect of LTV when using low-dose ATG remain to be elucidated. [Methods] We retrospectively analyzed 222 recipients who received allo-PBSCT using PTCY or low-dose ATG at Hokkaido University Hospital from January 2013 to July 2021. Of the 121 patients who underwent PBSCT with PTCY, 69 (all patients underwent Haplo-PBSCT) were without LTV prophylaxis and 52 (34 patients, Haplo-PBSCT) were with LTV. On the other hand, of the 101 patients who underwent PBSCT with low-dose ATG, 66 (12 patients, Haplo-PBSCT) were without LTV and 35 (2 patients, Haplo-PBSCT) were with LTV. LTV was administered on the day 0 at a dosage of 480 mg daily. All patients were monitored for CMV reactivation by using the anti-CMV pp65 monoclonal antibody HRP-C7 assay at least weekly from neutrophil engraftment to discharge and after discharge monthly until day 180 after SCT. CMV reactivation was defined as the start of CMV preemptive therapy, generally initiated when there are 2 or more CMV antigen positive cells per 50000 white blood cells. Moreover, CMV disease was defined by organ dysfunction attributable to CMV. [Results] Baseline characteristics of the patients are summarized in Table 1. Although more patients in ATG group compared to PTCY group received myeloablative conditioning regimen significantly, there was no significant difference between the groups in terms of sex, age, underlying disease, disease risk at SCT, and CMV serostatus. GVHD prophylaxis in PTCY consisted of CY (40-50 mg/kg on day 3 and 4), tacrolimus (from day 5), and mycophenolate mofetil (from day 5), whereas that in ATG consisted of ATG at a total dose of 2-3 mg/kg around day -2, tacrolimus (from day -1) and short-term methotrexate. LTV prophylaxis significantly reduced the cumulative incidence of CMV reactivation at 180 days after SCT in PTCY group (69.6% without LTV vs. 22.4% with LTV, p&amp;lt;0.001) and ATG group (62.6% without LTV vs. 18.9% with LTV, p&amp;lt;0.001; Figure 1.). Importantly, although CMV disease were occurred in 2 patients in PTCY (1 gastritis and 1 retinitis) and 8 focuses in 7 patients (3 pneumoniae, 3 enteritis, 1 retinitis, and 1 glossitis) in ATG without LTV prophylaxis, none of the patients receiving LTV developed CMV disease in both group. Moreover, LTV did not affect overall survival, the cumulative incidence of non-relapse mortality, relapse, acute GVHD, and engraftment of neutrophil and platelet. Interestingly, the cases of early CMV reactivation within 100 days after SCT under LTV prophylaxis was significantly more frequent in ATG than in PTCY (p=0.0345). [Conclusion] CMV reactivation occurred with similar frequency after allo-PBSCT with low-dose ATG as after that with PTCY. LTV was effective for prevention of CMV reactivation in both GVHD prophylaxis methods. CMV breakthrough reactivation may be likely to occur in PBSCT with low-dose ATG compared to that with PTCY even under LTV prophylaxis, and careful CMV monitoring may be required.

Cytomegalovirus-Seronegative Plus Leukoreduced Blood Products Are Still Superior to Only Leukoreduced Products in Preventing Transfusion-Transmitted Cytomegalovirus Infection

Introduction Cytomegalovirus (CMV) is one of the most important pathogens causing complications after hematopoietic stem cell transplantation (HSCT). Transfusion-transmitted CMV infection (TT-CMV) in CMV-seronegative patients is a major issue. Leukoreduced (LR) blood products were unavailable before 2004. However, they have gradually become widespread, and all blood products in Japan have been LR since 2007. This study aimed to assess the incidence of TT-CMV in CMV-seronegative HSCT recipients undergoing autologous HSCT (auto-HSCT) or allogeneic HSCT (allo-HSCT) from CMV-seronegative stem cell donors, who received transfusions with “CMV-seronegative products” from CMV-seronegative donors or “random donor products” from donors not tested for anti-CMV antibodies during the periods of availability and unavailability of LR products. Methods We retrospectively evaluated 3590 CMV-seronegative recipients registered in the TRUMP database of the Japanese Society for Transplantation and Cellular Therapy from 1999 to 2018. The recipients undergoing HSCT were categorized into three periods: Period A (when LR products were unavailable), 1999-2003 (n = 136); Period B (when there were both non-LR and LR products), 2004-2006 (n = 200); and Period C (when only LR products were available), 2007-2018 (n = 3254). TT-CMV was defined by the presence of CMV antigenemia and/or CMV diseases. This study was approved by the Ethics Committee of Kobe University Hospital and was conducted in accordance with the principles of the Declaration of Helsinki. Results The median age of the patients at HSCT was 40 years (range, 0-79 years). Among auto-HSCT patients (n = 1628) transfused with “CMV-negative products”(CN group) and “random donor products” (RD group), the incidences of TT-CMV in the CN and RD groups were 0% (0/4) and 3.1% (1/32) during Period A (P = 1.0), 22.2% (2/9) and 10.6% (7/66) during Period B (P = 0.29), and 3.8% (4/105) and 5.0% (71/1412) during Period C (P = 0.81), respectively. Among the allo-HSCT patients (n = 1962), the incidences of TT-CMV in the CN and RD groups were 10.0% (3/30) and 27.1% (19/70) during Period A (P = 0.07), 17.9% (5/28) and 29.9% (29/97) during Period B (P = 0.24), and 11.6% (41/355) and 28.0% (387/1382) during Period C (P &amp;lt; 0.001), respectively. During Period C, 99 patients in the CN group always underwent transfusion with leukocyte reduction filters, of which 11 (11.1%) were diagnosed with TT-CMV. In contrast, 202 patients in the CN group always underwent transfusion without the filters, of which 26 (12.9%) were diagnosed with TT-CMV (P = 0.78). In the RD group, 494 patients always underwent transfusion with leukocyte reduction filters, of which 124 (25.1%) were diagnosed with TT-CMV. On the other hand, 807 patients in the RD group always underwent transfusion without the filters, of which 245 (30.4%) were diagnosed with TT-CMV (P = 0.10). Multivariate analysis revealed that random donor products (hazard ratio [HR]: 3.11, 95% confidence interval [CI]: 2.13-4.52; P &amp;lt; 0.001) and age ≥16 years (HR: 2.90, 95% CI: 2.06-4.09; P &amp;lt; 0.001) were associated with an increased risk of TT-CMV in allo-HSCT. No significant difference was observed in the incidence of CMV diseases between the CN group (3.7%; 13/355) and the RD group (3.8%; 52/1382) (P = 1.00). The major CMV diseases included CMV colitis (n = 5) and pneumonia (n = 2) in the CN group, and CMV colitis (n = 30), pneumonia (n = 9), and retinitis (n = 4) in the RD group. Interestingly, among the patients with TT-CMV during Period C, 27/41 patients (65.9%) in the CN group and 333/387 patients (86.1%) in the RD group received early intervention with anti-CMV agents (P = 0.002). The limitation included that complement-fixation test for anti-CMV antibody was used in Japan Marrow Donor Program in this cohort. Conclusion This is the largest study demonstrating that CMV-seronegative plus LR products are still superior to only LR products from random donors in terms of TT-CMV prevention among allo-HSCT CMV-seronegative recipients. However, no significant difference was observed in the incidence of CMV diseases, possibly because of early intervention with anti-CMV agents.

IgG-antibodies to individual cytomegalovirus proteins and the peripheral blood lymphocyte subset profile in patients with anterior uveitis of varying severity

Background. Cytomegalovirus (CMV), a human beta-herpesvirus, persists latently lifelong after infection. CMV reactivation that occurs periodically can cause disease in target organs including the eye, not only in immunosuppressed, but also in immunocompetent people. Due to its immunosuppressive properties, persistent CMV is involved in the pathogenesis of diverse diseases. The aim was to study the features of antibody formation against six individual CMV proteins and to conduct a correlation analysis with parameters of peripheral blood lymphocyte profile in patients with anterior uveitis of varying severity (without hypopion-mild course, with hypopion-more severe course). Materials and methods. 36 patients with anterior uveitis were examined. Group I consisted of 20 patients without hypopion, group II 16 people with hypopion. No characteristic features of CMV anterior uveitis (elevated intraocular pressure, stromal iris atrophy) were found. Blood serum anti-CMV IgG antibodies (markers of chronic infection) were measured by ELISA, antibodies against individual recombinant CMV antigens were assessed by Line-Immunoassay specific to the main non-structural immediate early protein (IE), DNA-binding phosphoprotein p52, phosphoproteins of the tegument p150, p65, p28, GB-AD envelope glycoprotein. Lymphocyte subset composition was studied by flow cytometry: T-lymphocytes (CD3+), T-helper cells (CD3+CD4+CD8), T-cytotoxic (CD3+CD4CD8+), T-double-positive (CD3+CD4+CD8+), natural killers (CD16+CD56+), B-lymphocytes (CD19+). Statistical analysis was performed using StatTech v. 3.0.2 program (Stattech LLC, Russia). Results. In mild anterior uveitis, there was observed a significantly increased percentage of CD16+CD56+ and a decreased absolute number of CD3+CD8+ compared with anterior uveitis of a more severe course. Only in the group of patients with hypopion (more severe course) there was a direct significant correlation between level of antibodies against IE, p65, p28 antigens and percentage of double-positive cells (CD3+CD4+CD8+) as well as an negative significant relationship between the level of antibodies against IE, p65 and p52 antigens and CD19+ lymphocytes. Conclusion. The data obtained are consistent with few reports on the role of double-positive lymphocytes in the pathogenesis of severe forms of some viral diseases. Further studies are needed to assess an effect of individual viral antigens on CD3+CD4+CD8+ level in patients with chronic CMV infection to confirm the role of double-positive lymphocytes in the pathogenesis of a more severe course of anterior uveitis.

The impact of IdeS (imlifidase) on allo-specific, xeno-reactive, and protective antibodies in a sensitized rhesus macaque model.

Highly sensitized patients face many barriers to kidney transplantation, including higher rates of antibody-mediated rejection after HLA-incompatible transplant. IdeS, an endopeptidase that cleaves IgG nonspecifically, has been trialed as desensitization prior to kidney transplant, and successfully cleaves donor-specific antibody (DSA), albeit with rebound. IdeS was generated and tested (2mg/kg, IV) in two naïve and four allosensitized nonhuman primates (NHP). Peripheral blood samples were collected at regular intervals following IdeS administration. Total IgG, total IgM, and anti-CMV antibodies were quantified with ELISA, and donor-specific antibody (DSA) and anti-pig antibodies were evaluated using flow cytometric crossmatch. B cell populations were assessed using flow cytometry. IdeS successfully cleaved rhesus IgG in vitro. In allosensitized NHP, robust reduction of total, DSA, anti-pig, and anti-CMV IgG was observed within one day following IdeS administration. Rapid rebound of all IgG antibody populations was observed, with antibody levels returning to baseline around day 14 post-infusion. Total IgM level was not affected by IdeS. Interestingly, a comparable reduction in antibody populations was observed after the second dose of IdeS. However, we have not observed any significant modulation of B cell subpopulations after IdeS. This study evaluated efficacy of IdeS in the allosensitized NHP in IgG with various specificities, mirroring antibody kinetics in human patients. The efficacy of IdeS on preexisting anti-pig antibodies may be useful in clinical xenotransplantation. However, given the limitation of IdeS on its durability as a monotherapy, optimization of IdeS with other agents targeting the humoral response is further needed.

Assessment of mimicking by EBV-CMV immunoglobulin M of anti-HLA antibodies

ObjectiveWe aimed to show the cross-reactivity that may occur between immunoglobulin (Ig) M antibodies that form against Cytomegalovirus (CMV) and/or Epstein-Barr virus (EBV) and human leukocyte antigens (HLA). MethodsComplement-dependent cytotoxicity (CDC) cross-reactivity between serum samples of 57 patients with IgM positive CMV and/or EBV infections and T and B cells from 15 healthy donors were evaluated. Dithiothreitol was used to distinguish cross-reactivity caused by IgM antibodies from IgG. ResultsThe cross-reactivity ratio between pathogenic IgM antibodies with T cell of the 12th donor, and B cell of the 3rd, 4th, and 8th donors was significantly higher (p = 0.011, <0.001, <0.001 and 0.013, respectively). The ratio of B cell CDC cross-reactivity of all donors (26.4%) was higher than the ratio of T cell CDC cross-reactivity (5.2%) (p < 0.001). The ratio of T cell CDC cross-reactivity of sera containing both anti-CMV IgM and anti-EBV IgM antibodies was significantly higher than those of sera containing only anti-CMV IgM or only anti-EBV IgM antibodies (p = 0.002 and p < 0.001, respectively). There was no difference between B cell CDC cross-reactivity rates according to the presence of anti-CMV and/or anti-EBV IgM antibodies. ConclusionCross-reactivity may occur between anti-CMV and anti-EBV IgM antibodies with HLA molecules. Thus, in graft recipients, pathogenic IgMs can also act as de novo anti-HLA antibodies and aggravate the rejection process.

Cytomegalovirus and Epstein–Barr virus co-infected young and middle-aged adults can have an aging-related T-cell phenotype

Cytomegalovirus (CMV) is known to alter circulating effector memory or re-expressing CD45RA+ (TemRA) T-cell numbers, but whether Epstein–Barr virus (EBV) does the same or this is amplified during a CMV and EBV co-infection is unclear. Immune cell numbers in blood of children and young, middle-aged, and senior adults (n = 336) were determined with flow cytometry, and additional multivariate linear regression, intra-group correlation, and cluster analyses were performed. Compared to non-infected controls, CMV-seropositive individuals from all age groups had more immune cell variance, and CMV+ EBV− senior adults had more late-differentiated CD4+ and CD8+ TemRA and CD4+ effector memory T-cells. EBV-seropositive children and young adults had a more equal immune cell composition than non-infected controls, and CMV− EBV+ senior adults had more intermediate/late-differentiated CD4+ TemRA and effector memory T-cells than non-infected controls. CMV and EBV co-infected young and middle-aged adults with an elevated BMI and anti-CMV antibody levels had a similar immune cell composition as senior adults, and CMV+ EBV+ middle-aged adults had more late-differentiated CD8+ TemRA, effector memory, and HLA-DR+ CD38− T-cells than CMV+ EBV− controls. This study identified changes in T-cell numbers in CMV- or EBV-seropositive individuals and that some CMV and EBV co-infected young and middle-aged adults had an aging-related T-cell phenotype.

Cytomegalovirus antibodies are associated with mood disorders, suicide, markers of neuroinflammation, and microglia activation in postmortem brain samples.

Cytomegalovirus (CMV) is a common, neurotrophic herpesvirus that can be reactivated by inflammation and cause central nervous system disease. We hypothesize that CMV may contribute to the neuroinflammation that underlies some psychiatric disorders by (1) exacerbating inflammation through the induction of anti-viral immune responses, and (2) translating peripheral inflammation into neuroinflammation. We investigated whether the presence of anti-CMV antibodies in blood were associated with mental illness, suicide, neuroinflammation, and microglial density in the dorsolateral prefrontal cortex (DLPFC) in postmortem samples. Data (n = 114 with schizophrenia; n = 78 with bipolar disorder; n = 87 with depression; n = 85 controls) were obtained from the Stanley Medical Research Institute. DLPFC gene expression data from a subset of 82 samples were categorized into "high" (n = 30), and "low" (n = 52) inflammation groups based on a recursive two-step cluster analysis using expression data for four inflammation-related genes. Measurements of the ratio of non-ramified to ramified microglia, a proxy of microglial activation, were available for a subset of 49 samples. All analyses controlled for age, sex, and ethnicity, as well as postmortem interval, and pH for gene expression and microglial outcomes. CMV seropositivity significantly increased the odds of a mood disorder diagnosis (bipolar disorder: OR = 2.45; major depression: OR = 3.70) and among the psychiatric samples, of suicide (OR = 2.09). Samples in the upper tercile of anti-CMV antibody titers were more likely to be members of the "high" inflammation group (OR = 4.41, an effect driven by schizophrenia and bipolar disorder samples). CMV positive samples also showed an increased ratio of non-ramified to ramified microglia in layer I of the DLPFC (Cohen's d = 0.81) as well as a non-significant increase in this ratio for the DLPFC as a whole (d = 0.56). The results raise the possibility that the reactivation of CMV contributes to the neuroinflammation that underlies some cases of psychiatric disorders.

Coordination of NK cell markers expression and IgG response in hCMV infection

Human cytomegalovirus (hCMV) is a prevalent virus that affects a large proportion of the population worldwide. Natural Killer (NK) cells are essential immune cells that play a crucial role in controlling hCMV infection. Despite the wide spread of hCMV infection, there is still not enough data related to the association between innate and adaptive immunity. This study investigated the coordination between some of the NK cell markers expression and humoral immune response during hCMV infection. Thirty-three samples obtained from different healthy donors were investigated. The anti-hCMV IgG antibody titer was measured in serum samples, and expression of NKG2C, HLA-DR, CD57, KIR2DL2/DL3, and KIR2DL1 were analyzed in CD56+CD3- cells in PBMC samples by flow cytometry. To evaluate the dependence of proportions of different NK cell subsets on IgG titers, cluster analysis was first performed on all the obtained data, resulting in the identification of four main clusters. The identified clusters demonstrated a dependence on the levels of hCMV antibodies, according to which clusters corresponding to seronegative and low-positive were grouped. The results confirmed that hCMV infection leads to an expansion of NK cell populations expressing the NKG2C marker, which correlates with higher levels of IgG response to hCMV. Besides, we identified increased HLA-DR+ and decreased of KIR2DL1+ NK cells proportions in the middle anti-CMV-IgG level group compared to samples obtained from seronegative and low-positive donors. Moreover, the statistically significant negative correlation was found between KIR2DL1+NK cell percentage and anti-CMV IgG antibody titer, while the positive correlation between HLA-DR+NK cell proportion and the IgG level was noticed only without the cluster corresponded to high level of anti-hCMV IgG. In this cohort, we did not find any association between KIR2DL3 and CD57 expression in NK cells and levels of IgG response to hCMV. This may indicate that different subsets of NK cells may have distinct roles in regulating humoral immunity to hCMV. Overall, the results of the study provide valuable insights into the coordination of NK cell marker expression and IgG response in hCMV infection.

Impact of Cytomegalovirus Infection and Disease on Graft Loss After Pancreas Transplantation: A Single-Institution Study in Japan

Cytomegalovirus (CMV) is one of the most frequent infections after pancreas transplantation (PTx), and it is unclear whether CMV infection is associated with pancreas graft loss. A limited number of studies about the relationship between CMV infection and pancreas graft loss have been reported from Western countries, but there have been no reports from Japan. This study investigated the relationship between CMV infection and pancreas graft loss after PTx in a single Japanese institution. This study included 58 patients who underwent PTx from deceased donors from April 2000 to March 2021 in our institution. We assessed pancreas graft loss based on CMV infection and disease and investigated the causes of graft loss, the time of onset of CMV disease, and the time of graft loss for each case. The numbers of patients in the 4 categories of donor (D) and recipient (R) pretransplant anti-CMV antibody status were as follows: 4 (6.9%) in the D-/R- group, 6 (10.3%) in the D-/R+ group, 34 (58.6%) in the D+/R+ group, and 14 (24.1%) in the D+/R- group. Of the 58 patients, 74.1% and 44.1% received diagnoses of CMV infection and disease after PTx, respectively. There were no significant differences in the survival rates of pancreas graft loss stratified by CMV infection (P=.1809) or disease (P=.6241). This study suggests that CMV infection and disease had no significant influence on pancreas graft loss in this Japanese institution.

Acquisition of Antibody Against Cytomegalovirus After Kidney Transplantation in Seronegative Recipients

Cytomegalovirus (CMV) infection is one of the most important infectious diseases affecting recipients of kidney transplantation (KTx). However, the timing of seroconversion for CMV infection in seronegative recipients remains unclear. We evaluated CMV infections in CMV-seronegative recipients and the time to acquire antibodies against CMV. We conducted a retrospective study of 228 recipients who underwent KTx between March 1988 and February 2018 at the Niigata University Hospital. The anti-CMV IgG antibody profile before and after transplantation was analyzed. Oral acyclovir or valganciclovir was used as prophylactic therapy for at least 6 months after transplantation. Cytomegalovirus infection was defined as CMV viremia detected using the CMV pp65 antigenemia assay. In this study, 50 cases (21.9%) were CMV-seronegative recipients. Over a median follow-up period of 126.7 months, 68% (34/50) of CMV-seronegative recipients experienced CMV viremia or overt disease symptoms as the first episode of CMV infection. The median duration from transplant to CMV infection was 69.0 days (range, 22-7426). All the recipients who experienced CMV infections acquired seroconversion. The median duration from KTx to seroconversion was 7.2 months (range, 2.8-252.3). Almost all CMV-seronegative recipients could acquire anti-CMV IgG antibodies within 2.5 years. In seronegative recipients whose donors were seronegative, no CMV viremia was found, and none developed anti-CMV IgG antibodies. In the clinical practice of CMV-seronegative recipients, we should consider that physicians must closely monitor the occurrence of CMV infection up until 2.5 years after KTx.

Prevalence of Cytomegalovirus infection among pregnant women with cord blood examination

Objective: This study aimed at determining the prevalence of CMV infection among pregnant women at the end of pregnancy and CMV transmission to their newborns.&#x0D; Methods: This is cross sectional study, 213 pregnant women at delivery and their newborn babies from the Obstetrics and Gynecology Hospital in Duhok /Iraq were enrolled. A questionnaire was prepared to be answered by participants, including age, place of residence, educational level, and obstetric history as number of births, any bad obstetric history such as abortion, still birth, intrauterine growth retardation, congenital anomalies after birth.3-5 ml of blood was drawn from each woman and examined by ELISA kit to check for the presence of Anti CMV IgM, IgG, then IgG avidity test for those with positive (IgG and IgM). Samples of cord blood were collected from newborns after birth and checked for the presence of CMV IgM by ELISA and CMV –DNA by conventional PCR using specific primers to diagnose congenital infection and determine the rate of viral transmission from infected women.&#x0D; Results: Serological examinations showed that 212 (99.5%) participants were CMV-IgG positive, 15 (7%) were positive for anti-CMV IgM and IgG antibodies, IgG avidity test for 15 women were of high avidity (&gt;89%) which indicated non primary infections. Cord blood of newborns of those 15 women with positive IgG and IgM tested negative for Anti CMV IgM by ELISA and no CMV-DNA was detected by PCR, which revealed no transmission from those pregnant to their newborns.&#x0D; Conclusion: This study demonstrated high prevalence of CMV among examined pregnant women in Duhok city which makes them prone to non-primary infection. IgG avidity test is of high efficacy to interpret the detection of IgG and IgM together in pregnant women. Cord blood examination for the existence of CMV-IgM and CMV-DNA after delivery could exclude congenital infection.

Efficiency of CMV serodiagnosis during pregnancy in daily laboratory routine

BackgroundMaternal acute primary cytomegalovirus (CMV) infection during the first trimester may cause severe long-term sequelae in newborns. For risk assessment, serological screening is routinely performed in pregnant women based on IgM, IgG and avidity tests using whole-virus antigen. A recent study evaluated the diagnostic value of recombinant protein-based ELISAs as second-line tests in pregnancy CMV screening, including anti-p52 IgM and anti-gB IgG as markers defining the early and late phase of infection, respectively. In the present study, these recombinant ELISAs were used as first-line screening tests in daily laboratory routine and compared to lysate-based assays with respect to [i] the number of conclusive results obtained with the initial sample and [ii] the underlying workload. Methods553 unselected routine serum samples from pregnant women were tested for anti-CMV IgM and IgG antibodies using lysate-based ELISAs and avidity testing. Anti-CMV IgM antibodies against recombinant p52 and anti-CMV IgG antibodies against recombinant glycoprotein B (gB) were also determined by ELISA. All assays were performed and interpreted according to the manufacturer’s instructions. ResultsFor lysate-based IgM, IgG and avidity testing, 84.6 % of samples yielded conclusive results in a total of 1156 tests, while 15.4 % needed follow-up testing of a consecutive sample. Anti-p52 CMV IgM and anti-gB CMV IgG testing produced conclusive results for 92.8 % of samples in a total of 1026 tests, while 7.2 % samples required follow-up testing. ConclusionsThe first-line use of ELISAs measuring anti-p52 CMV IgM and anti-gB CMV IgG antibodies to test for maternal CMV infection increases the number of conclusive results derived from an initial serum sample while requiring a considerably lower number of tests compared to the lysate-based approach. For day-to-day routines in a diagnostic laboratory, this high efficiency of the recombinant testing approach has significant practical relevance.