Cellular Immune Deficiency Research Articles

Diagnostic Modalities in Primary Immunodeficiency.

As the field of inborn errors of immunity expands, providers continually update and fine-tune their diagnostic approach and selection of testing modalities to increase diagnostic accuracy. Here, we first describe a mechanistic consideration of laboratory testing, highlighting both benefits and drawbacks of currently clinically available testing modalities. Next, we provide methods in evaluation of patients presenting with concern for inborn errors of immunity as defined by the International Union of Immunological Societies 2019 phenotypic categories: primary antibody deficiencies, cellular and humoral immune deficiency, disorders of the innate immune system, and syndrome-associated and primary immune regulation disorders (PIRDs). Using the suggested approach in this paper as a roadmap highlights the importance of thorough history taking and physical examination as the foundation to guide further diagnostic tests. This is followed by enumeration and functional testing. Finally, to determine the underlying molecular etiology-specific genetic panels, chromosomal microarrays, and broad genetic testing (whole exome sequencing or whole genome sequencing) are available.

Investigating the Relation between LCK Gene Expression with Type 2 Diabetes Patients in Yazd Diabetes Research Center

Objective: Type 2 diabetes mellitus (T2DM) is characterized by insulin resistance and insulin secretory defect. Deficiency of cellular immunity is known as one of the factors involved in the pathogenesis of T2DM. lymphocyte-specific protein tyrosine kinase( LCK) is an important gene involved in the intracellular signaling pathways of lymphocytes. This study aimed at determining and comparing LCK gene expression levels in diabetic patients compared with the healthy controls. Materials and Methods: In this case-control study, 60 people, including 30 T2DM and 30 healthy people were included. The expression levels of the LCK gene were measured by real-time polymerase chain reaction and the obtained data were analyzed by T-test in GraphPad Prism6 software. Results: The expression level of the LCK gene was increased in diabetic samples compared with the healthy samples (P= 0.0001). Conclusion: The results suggested that changes in the expression levels of LCK gene can play a role in the pathogenesis of T2DM.

Pathomorphology of lymph nodes in patients with severe SARS-CoV-2 infection observed in Primorsky Krai

Introduction. When studying COVID-19 pathology, considerable attention is paid to the damage to the respiratory and cardiovascular systems, which are associated with the manifestation of the infection. Data on changes in the organs of the lymphatic system are yet scarce. To date, COVID-19 is sure to cause the dysfunction of the immune system; however, information about the damage to the lymph nodes is ambiguous. The aim of the study was to characterize morphological changes in the hilar lymph nodes of patients who died from a new coronavirus infection COVID-19 in Vladivostok in 2021. Materials and methods. A morphological study of the lymph nodes was performed in 20 patients who died from the new coronavirus infection, diagnosed in vivo using a PCR test. The controls were patients who did not have diseases of the hematopoietic and lymphoid tissue in their medical histories but who died a violent death. The material for histological examination was processed according to generally accepted methods. Results. In all patients, we identified lymphadenopathy with hypoplasia of lymphoid tissue of varying severity. In the cortex, lymphatic follicles were detected, mainly without light (germinal) centers, as well as follicles with a pronounced rarefaction of cells and exposure to the reticular stroma, with no visualization of the paracortical zone. According to the severity of reactive changes in the T- and B-dependent zones of the lymph nodes, we distinguished two types of lymphadenopathy: 1) mixed follicular type; 2) follicular involution with lymphoid depletion. Conclusion. Pathological changes in the hilar lymph nodes of the lungs in patients who died from COVID-19 indicated immunosuppressive effects of the SARS-CoV-2. The pathologic changes in lymph nodes manifested with lymphocytic depletion in T- and B-dependent zones. This indicates a deficiency of cellular and humoral immunity in moderate and severe COVID-19. Keywords: coronavirus infection, COVID-19, pathomorphology, lymph nodes, lymphocytic depletion, immunodeficiency

Regulatory T-Cells and Multiple Myeloma: Implications in Tumor Immune Biology and Treatment.

Multiple myeloma (MM) is associated with both cellular and humoral immune deficiencies and, despite significant advances in treatment, remains an incurable disease. Regulatory T-cells (Tregs) represent a critical subset of CD4 T-cells, characterized by CD4 + CD25+ Forkhead box P3+ (FoxP3+) phenotype, able to control peripheral tolerance and responses to foreign and tumor antigens. Tregs are elevated in various types of cancer, including hematological malignancies; in MM, data regarding Tregs function and numbers and their correlation with survival parameters are controversial. Advances in cancer biology have shown that the tumor microenvironment plays an important role in tumor progression. In MM, the highly immunosuppressive nature of the bone marrow microenvironment has been significantly elucidated in the past decade and it is now well acknowledged that targeting only the tumor clone may not be able to cure MM. Tregs within the tumor microenvironment might play a significant role in the suppression of antitumor immune responses against cancer cells and are considered to predict poor outcome in cancer patients; nonetheless the exact prognostic significance of this cell subpopulation in malignancies is still a matter of debate. In this review, we discuss the role of Tregs as an essential cell population of the MM immune microenvironment.

Could β-Lactam Antibiotics Block Humoral Immunity?

It has been reported that treatment with β-lactam antibiotics induces leukopenia and candidemia, worsens the clinical response to anticancer immunotherapy and decreases immune response to vaccination. β-lactamases can cleave β-lactam antibiotics by blocking their activity. Two distincts superfamilies of β-lactamases are described, the serine β-lactamases and the zinc ion dependent metallo-β-lactamases. In human, 18 metallo-β-lactamases encoding genes (hMBLs) have been identified. While the physiological role of most of them remains unknown, it is well established that the SNM1A, B and C proteins are involved in DNA repair. The SNM1C/Artemis protein is precisely associated in the V(D)J segments rearrangement, that leads to immunoglobulin (Ig) and T-cell receptor variable regions, which have a crucial role in the immune response. Thus in humans, SNM1C/Artemis mutation is associated with severe combined immunodeficiency characterized by hypogammaglobulinemia deficient cellular immunity and opportunistic infections. While catalytic site of hMBLs and especially that of the SNM1 family is highly conserved, in vitro studies showed that some β-lactam antibiotics, and precisely third generation of cephalosporin and ampicillin, inhibit the metallo-β-lactamase proteins SNM1A & B and the SNM1C/Artemis protein complex. By analogy, the question arises as to whether β-lactam antibiotics can block the SNM1C/Artemis protein in humans inducing transient immunodeficiency. We reviewed here the literature data supporting this hypothesis based on in silico, in vitro and in vivo evidences. Understanding the impact of β-lactam antibiotics on the immune cell will offer new therapeutic clues and new clinical approaches in oncology, immunology, and infectious diseases.

Clinical findings in patients with leprosy who are infected with COVID-19: a case series from Elazığ, Turkey

Background: Leprosy is a chronic granulomatous infectious disease caused by Mycobacterium leprae that can be self-limiting or progressive according to the immunological status of the host organism. Aim and objectives: We present the demographic and clinical characteristics of lepromatous leprosy (LL) patients who had Covid-19 infection. Materials and methods: This study recruited patients hospitalized in Elazığ Leprosy Hospital. Details of the patients were collected as follows: age, gender, presence of comorbidity, leprosy classification, clinical presentation, presence of leprosy reactions and complaints at the time of admission; laboratory and computed tomography (CT) findings and treatments applied. Results: This study included 9 patients with lepromatous leprosy (LL classification). Six patients were PCR positive for SARS-CoV-2, 7 patients had thoracic CT involvement. The mean age was 78.7 years (range 65–95). Complaints on admission included fatigue (n:7), cough (n:4) and fever (n:3) followed by sore throat (n:1), headache (n:1) and dysphonia (n:1). Two patients were asymptomatic, PCR positivity was detected in one and CT positivity was detected in another during screening. All patients received treatment with favipiravir and enoxaparin. No leprosy reactions occurred. All patients were discharged with recovery after completion of treatment. Conclusion: We expected that the risk for catching COVID-19 may be higher in LL patients who have a cellular immunity deficiency, and that the course of the infection may be more severe in COVID-19-infected LL patients, for the same reason. However, we have seen a decrease in the clinical severity of the COVID-19 and no patient losses. We conclude that this situation may be due to the drugs used for leprosy treatment. These immune-modulating drugs may cause a decrease in the clinical severity of the COVID-19, but we need larger studies on this subject.

Deep Dermatophytosis

Deep dermatophytosis is a rare fungal infection described for the first time by Hadida and Schousboe. It is a chronic dermatophytic infection of the skin and viscera, mainly described in the Maghreb where it has an autosomal recessive transmission. Immunological studies have revealed a deficiency of cellular immunity leading to dermatophytes tolerance. Commonly, first lesions begin in childhood. Both superficial and deep dermatophytic lesions are possible. A 17 years old male patient with a history of chronic generalized pruritic erythematous-squamous lesions presented for subcutaneous abscesses of scalp and trunk. He underwent multiple antibiotic courses and surgical excisions with persistent relapses. Trichophyton rubrum was identified in skin biopsy and all samples sites. It was found no immunodeficiency within the limits of the blood tests conducted. Search for CARD9 protein deficiency was positive. Treatment based on Terbinafine and surgical drainage with mid improvement

Иммунологические показатели как прогностические маркеры исхода суставсберегающих операций у детей с ишемическими деформациями проксимального отдела бедра (предварительные результаты)

Objective Assess the immune status of children with ischemic deformity of the proximal femur taking into account the treatment outcome. Material and methods Preoperative assessment of the immune status was performed for 16 adolescents with ischemic deformity of the proximal femur. Patients' age was 14.4 ± 0.45 years. Immunological parameters of 12 adolescents with satisfactory (group I) and 4 patients with poor (group II) anatomical and functional outcome of surgical treatment were compared. Peripheral blood lymphocyte typing was produced with laser cytometry assay, and IgA, IgM, IgG, IgE, IL-18, IFNγ and circulating immune complexes were quantified with enzyme-linked immunoassay. Results Сomparative analysis of preoperative immunological parameters showed statistically significant decline of Т-helper cells (СD3+СD4+), high levels of activated СD3+CD25 and СD3+HLADR T lymphocytes, low levels of activated NК cells (СD8dimCD38+) and IFNγ in group II. Humoral immune parameters indicated to low levels of IgA, IgM, IgG and higher levels of circulating immune complexes in group II relative to group I. Conclusion Preliminary data obtained suggest that cellular and humoral immune deficiency can be associated with unfavorable outcome of joint preservation procedure. Such immunological parameters as T lymphocytes subpopulation, immunoglobulins, circulating immune complexes and IFNγ can be used as significant prognostic factors for preoperative planning and postoperative rehabilitation of adolescents with ischemic deformity of the proximal femur.

Sars Cov-2/COVID-19 in Multiple Myeloma Latin-American Patients COVID-Lamm Study on Behalf of Gelamm (Grupo de Estudio Latino- Americano de Mieloma Múltiple)

BackgroundThe SARS CoV-2 / COVID 19 pandemic has challenged the world's health systems, especially services that treat cancer. The first studies in China showed that cancer patients had a higher risk of becoming infected and dying. Other risk factors for mortality were age over 65 years, male sex, the presence of comorbidity, hypertension, cardiovascular disease, diabetes, and obesity. Data on the specific behavior of patients with multiple myeloma (MM) in the pandemic are scarce. The mechanisms by which MM patients may have a higher mortality are multiple, derived both from the disease itself due to cellular and humoral immunity deficiency as well as from anti-myeloma treatment. The present study aims to establish the behavior of the disease in the pandemic period in Latin America.MethodsThis is a retrospective case series of in and outpatients with a diagnosis of MM and COVID-19 reported from centers from Latin America between March and July 31, 2020. The analysis of demographic, clinical, laboratory, complications and therapy variables were done using descriptive statistics. A Kaplan Meier survival analysis was performed, with Log Rank statistic. Finally, a Cox regression was performed to identify independent risk factors of worse outcome. Pre-admission characteristics, MM status, and comorbidities constituted the reference model and were used to adjust the association of relevant MM characteristics with mortality. Program used for analysis was SPSS statistics 25.ResultsFifty-two patients with COVID-19 and MM from 7 countries were included. Demographic characteristics, comorbidities, infection baseline clinical conditions, treatment, and outcomes are shown in Table 1. The characteristics in terms of MM status are shown in Table 2.When performing the survival analysis, it is evidenced that the survival of the entire cohort at day 49 was 67% Figure 1. When we focus on patients with comorbidities, survival drops to 53.5% +/- 10.6 (CI 95% 53.4 - 99.4 and p value of 0.041) for the same day Figure 2. When performing the obesity analysis, a drop in survival of up to 39% was observed (95% CI 24.448 - 56.76, with p = 0.00001) Figure 3. Adjusted HR for obesity is 5,078 (95% CI 1,389-18,558, α0.014) and mechanical ventilation with a HR of 3,943 (95% CI - 1,296 - 11,998, α0.016)When comparing patients with controlled MM (> PR) versus uncontrolled, the mortality rate was 84% versus 58% respectively (p = 0.109). Comorbidities (presence of either diabetes, arterial hypertension, cardiovascular disease, or “others”), obesity, need of mechanical ventilation, and <VGPR at the time of infection were independent factors of lower survival Table 3.ConclusionsPatients with MM and COVID-19 has an overall mortality of approximately 30% and this is strongly influenced by the presence of comorbidities, uncontrolled disease and need of mechanical ventilation. Survival of patients without comorbidities and controlled disease is good, suggesting that this group of patients would not require modification of MM therapy. The main limitations of our study are its retrospective nature and the low number of patients. It must be highlighted that at the time of the analysis most of Latin American countries were still in the peak of the pandemic. Prospective studies are required to elucidate the behavior of these risk factors in mortality, to optimize the management of patients with MM in this period of the SARS CoV-2 / COVID-19 pandemic.DisclosuresPeña:BindingSite: Research Funding; Janssen: Membership on an entity’s Board of Directors or advisory committees, Speakers Bureau; Sandoz: Membership on an entity’s Board of Directors or advisory committees; Roche: Membership on an entity’s Board of Directors or advisory committees; Abbvie: Membership on an entity’s Board of Directors or advisory committees; Amgen: Speakers Bureau. Abello:Novartis: Consultancy, Honoraria; Amgen: Consultancy, Research Funding; Takeda: Honoraria, Research Funding; Dr. Reddy’s: Consultancy, Research Funding; Abbvie: Consultancy, Research Funding. Idrobo:Abbvie: Honoraria, Speakers Bureau; Tecnofarma: Honoraria, Speakers Bureau; Takeda: Honoraria, Speakers Bureau; Janssen: Honoraria, Speakers Bureau; Amgen: Honoraria, Speakers Bureau. Rojas:Novartis: Consultancy; Roche: Honoraria; Sandoz: Honoraria; Abbvie: Honoraria. Remaggi:Takeda: Honoraria; Raffo Argentina: Honoraria, Research Funding; Janssen: Honoraria, Research Funding; Gador Argentina: Research Funding; Amgen: Honoraria, Research Funding. Alvarado:Celgene: Speakers Bureau; Alexion: Speakers Bureau; Amgen: Speakers Bureau; Novartis: Speakers Bureau; Roche: Speakers Bureau. De la Peña-Celaya:Amgen: Speakers Bureau; Janssen: Speakers Bureau; Novartis: Speakers Bureau. Perez:Novartis: Speakers Bureau; Celgene: Speakers Bureau; Roche: Speakers Bureau.

Epidemiological and clinical features of 201 COVID-19 patients in Changsha city, Hunan, China.

In December 2019, a cluster of coronavirus Disease 2019 (COVID-19) occurred in Wuhan, Hubei Province, China. The present study was conducted to report the clinical characteristics of 201 COVID-19 patients in Changsha, China, a city outside of Wuhan. All of the patients with confirmed COVID-19 were admitted to the First Hospital of Changsha City, the designated hospital for COVID-19 assigned by the Changsha City Government. The clinical and epidemiological characteristics, data of laboratory, radiological picture, treatment, and outcomes records of 201 COVID-19 patients were collected using electronic medical records. This study population consisted of 201 hospitalized patients with laboratory-confirmed COVID-19 in Changsha by April 28, 2020. The median age of the patients was 45 years (IQR 34–59). About half (50.7%) of the patients were male, and most of the infected patients were staff (96 [47.8%]). Concerning the epidemiologic history, the number of patients linked to Wuhan was 92 (45.8%). The most common symptoms were fever (125 [62.2%]), dry cough (118 [58.7%]), fatigue (65 [32.3%]), and pharyngalgia (31 [15.4%]). One hundred and forty-four (71.6%) enrolled patients showed bilateral pneumonia. Fifty-four (26.9%) patients showed unilateral involvement, and three (1.5%) patients showed no abnormal signs or symptoms. The laboratory findings differed significantly between the Intensive Care Unit (ICU) and non-ICU groups. Compared with non-ICU patients, ICU patients had depressed white blood cell (WBC), neutrocytes, lymphocytes, and prolonged prothrombin time (PT). Moreover, higher plasma levels of erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), procalcitonin (PCT), alanine aminotransferase (ALA), aspartate aminotransferase (AST), creatine kinase (CK), creatine kinase-MB (CK-MB), creatinine (CREA), and lactate dehydrogenase (LDH) were detected in the ICU group. In this single-center study of 201 COVID-19 patients in Changsha, China, 22.4% of patients were admitted to ICU. Based on our findings, we propose that the risk of cellular immune deficiency, hepatic injury, and kidney injury should be monitored. Previous reports focused on the clinical features of patients from Wuhan, China. With the global epidemic of COVID-19, we should pay more attention to the clinical and epidemiological characteristics of patients outside of Wuhan.

T Lymphocytes in Patients With Nijmegen Breakage Syndrome Demonstrate Features of Exhaustion and Senescence in Flow Cytometric Evaluation of Maturation Pathway.

Patients with Nijmegen Breakage Syndrome (NBS) suffer from recurrent infections due to humoral and cellular immune deficiency. Despite low number of T lymphocytes and their maturation defect, the clinical manifestations of cell-mediated deficiency are not as severe as in case of patients with other types of combined immune deficiencies and similar T cell lymphopenia. In this study, multicolor flow cytometry was used for evaluation of peripheral T lymphocyte maturation according to the currently known differentiation pathway, in 46 patients with genetically confirmed NBS and 46 sex and age-matched controls. Evaluation of differential expression of CD27, CD31, CD45RA, CD95, and CD197 revealed existence of cell subsets so far not described in NBS patients. Although recent thymic emigrants and naïve T lymphocyte cell populations were significantly lower, the generation of antigen-primed T cells was similar or even greater in NBS patients than in healthy controls. Moreover, the senescent and exhausted T cell populations defined by expression of CD57, KLRG1, and PD1 were more numerous than in healthy people. Although this hypothesis needs further investigations, such properties might be related to an increased susceptibility to malignancy and milder clinical course than expected in view of T cell lymphopenia in patients with NBS.

Modulatory effect of Tim-3/Galectin-9 axis on T-cell-mediated immunity in pulmonary tuberculosis

Patients affected by pulmonary tuberculosis (PTB) manifest deficiencies in innate cellular immunity. The Tim3/Galectin-9 axis is an important regulator of Th1 cell immunity, leading to Th1 cell apoptosis. Herein, this study aims to clarify the underlying roles of the Tim-3/Galectin-9 axis in T-cell immunity in PTB. Peripheral blood mononuclear cells (PBMCs) were extracted from subjects with and without PTB to examine the expression of CD4, CD8, CD25, and Tim-3 under the stimulation of Mycobacterium tuberculosis (MTB) and purified protein derivative (PPD). In addition, the expression of Tim-3 and Galectin-9 in PBMCs was determined. The Tim-3/Galectin-9 axis in the PBMCs was activated or blocked to detect the secreted levels of IFN-γ, TNF-α, IL-2, and IL-22. MTB stimulation increased the expression of CD4, CD8, CD25, Tim-3, and Galectin-9 in PBMCs. The blockade of Tim-3/Galectin-9 axis resulted in reduced secretion of IFN-γ, TNF-α, IL-2, and IL-22 from T-cells. Moreover, Tim-3+CD4+T, Tim-3+CD8+, and Tim-3+CD25+T cells exhibited a greater ability to inhibit the replication of MTB in macrophages. Taken conjointly, the blockade of Tim-3/ Galectin-9 axis inhibits the secretion of inflammatory cytokines in T-cells to regulate the T-cell immunity in PTB.

Pertinence of immune waning in measles virus infection – a short review

A striking occurrence will mark an acute infection with the measles virus. The patient soon acquires a lifelong specific immunity to the virus. But, unexpectedly, a transient cellular immune deficiency takes place as well, for several months. The causes of this paradox have not been entirely elucidated. An atypical measles syndrome was scrutinized in order to evaluate the immune status of patients who developed this disorder, subsequently to the administration of the formalin inactivated measles virus vaccine. Although considered not to occur in classic measles, an immune waning develops, at least, in atypical measles syndrome.

The pattern of mucocutaneous disorders in pediatric HIV patients in a tertiary care center

Background: Dermatological manifestations constitute one of the most common clinical features in HIV-infected children. An early recognition of such features is important for an early diagnosis and also to assess the prognosis of HIV infection. Objective: The present study was conducted to determine the prevalence and pattern of mucocutaneous manifestations in HIV-infected children and also to assess the association between those manifestations and degree of cellular immune deficiency. Materials and Methods: This was a cross-sectional study conducted in the Outpatient Department of SKIN and STD between April 2017 to March 2018. A total of 100 HIV patients

Latent Membrane Protein 1 Contributes to Deficient Cellular Immunity in NKTCL Patients

Introduction Extranodal NK/T cell lymphoma (NKTCL) is an aggressive malignance that is correlated closely with persistent Epstein-Barr virus (EBV) infection and belongs to a latency II EBV disease. Chronic persistent viral infection is known to result in T cell exhaustion, which will dramatically impede anti-tumor immunity. Delineating mechanisms of immunosuppression contributes to the development of novel immuno-therapeutically strategies. Herein, we described the immune status and possible role of latent membrane protein 1(LMP1) in NKTCL. Methods Peripheral blood mononuclear cells (PBMCs) from newly diagnosed NKTCL patients and age-matched healthy donors (HDs) were isolated and stained with surface marker or intracellular marker after permeabilization. The frequency of EBV-specific cytotoxic T cells (CTLs) was analyzed by staining with HLA-A2 tetramers assembled with synthetic peptides from LMP1. T cell proliferation was detected by dilution of stained CFSE, and apoptosis of T cells was performed via annexinV and 7-AAD dual-staining. Cytotoxicity assay of natural killer cells (NK) was measured by detecting apoptosis of CFSE-labeled K562 cells after co-culturing with PBMCs. All flow cytometry was performed by BD FACS CantoII, and analyzed with FlowJo software. Results PBMCs from 19 NKTCL patients and 18 HDs were analyzed, which showed that the patients had higher ratio of CD4+/CD8+ cells and lower frequency of CD3-CD56+NK cells, higher percentage of immunosuppressive CD4+CD25hiCD127low T regulatory cells (Tregs) and HLA-DR-CD11b+CD33+ myeloid derived suppressor cells (MDSCs). Notably, the ratio of CD8+/Tregs, a parameter representing immune effector cells status, was obviously decreased in NKTCL patients (Figure-a). Given the potential association of T cell exhaustion and NKTCL, we detected expression of T cell exhaustion markers on T cells from NKTCL patients, and found that both CD4+ and CD8+ T cells expressed much higher level of inhibitory molecules PD1, CTLA4, TIGIT and TIM3 in NKTCL patients than that of HDs (Figure-b). To further explore cellular immunity in NKTCL, T cells were divided into four subsets: CD45RA-CCR7+ central memory T cells (Tcm), CD45RA+CCR7+ terminally differentiated effector T cells (Temra), CD45RA+CCR7- naïve T cells (Tn), CD45RA-CCR7- effector memory T cells (Tem) (Figure-c). It showed that CD4+ T cells had lower Tcm, Temra and Tn but much higher proportion of Tem compared with HDs (Figure-d). Moreover, we found that expression of PD1 and CTLA4 were upregulated on all lymphocyte subsets in NKTCL patients than HDs (Figure-e). We collected additional 13 NKTCL patients and 10 HDs with positive HLA-A2 phenotype to measure percentages of EBV-specific CTLs, which showed that frequency of EBV-specific CTLs was remarkably decreased in NKTCL patients (Figure-f). In addition, EBV-specific CTLs from NKTCL patients were more likely to express higher levels of exhaustion markers but produced much less IFN-γ (Figure g-h). To explore the potent mechanism of immunosuppression in NKTCL, LMP1 attracted our attention, which had been proven capable of activating multiple signaling pathways to exert its oncogenesis function. To elucidate effect of LMP1 on immune cells, we constructed two LMP1-derived peptides and found that LMP1-derived peptides suppressed the proliferation of CD4+ and CD8+ T cells and inhibited IFN-γ production of CD8+ T cells and NK cells (Figure i-j). Meanwhile, LMP1 promoted apoptosis of both CD4+ and CD8+ T cells (Figure-k). Subsequently regulation of LMP1 on exhaustion markers of T cells were detected, which showed that PD1, CTLA4, TIGIT and TIM3 were significantly upregulated on both CD4+ and CD8+ T cells after treatment with LMP1 derived peptides (Figure-m). Furthermore, LMP1 impaired NK cytotoxicity ability, evidenced by decrease apoptosis of target cells (Figure-n). Besides its effect on immune effector cells, we also found that LMP1-expressing NKTCL cell lines obviously induced both Tregs and MDSCs after co-culture. Interestingly, the extent of induction by LMP1 of Tregs and MDSCs were in line with the expression level of LMP1 on NKTCL cells (Figure o-p). Conclusion Our study showed that LMP1 contributes to deficient cellular immunity in NKTCL patients, providing us with more insight into immunosuppressive in this entity of disease, which would lead to identification of novel treatment strategies. Figure Disclosures No relevant conflicts of interest to declare.

Flow Cytometric Determination of Actin Polymerization in Peripheral Blood Leukocytes Effectively Discriminate Patients With Homozygous Mutation in ARPC1B From Asymptomatic Carriers and Normal Controls.

Actin nucleators initiate formation of actin filaments. Among them, the Arp2/3 complex has the ability to form branched actin networks. This complex is regulated by members of the Wiscott-Aldrich syndrome protein (WASp) family. Polymerization of actin filaments can be evaluated through flow cytometry by fluorescent phalloidin staining before and after stimulation with N-formyl-methionyl-leucyl-phenylalanine (fMLP). We identified a missense mutation in the gene ARPC1B (Arp2/3 activator subunit) resulting in defective actin polymerization in four patients (three of them were related). All patients (1 male, 3 female) developed microthrombocytopenia, cellular immune deficiency, eczema, various autoimmune manifestations, recurrent skin abscesses and elevated IgE antibodies. Besides four patients with homozygous mutation in ARPC1B, we also identified six heterozygous carriers without clinical disease (3 males, 3 females) within the same family. We developed a functional test to evaluate Arp2/3 complex function, which consists of flow cytometric detection of intracellular polymerized actin after in vitro fMLP stimulation of leukocytes. Median fluorescence intensities of FITC-phalloidin stained actin were measured in monocytes, neutrophils and lymphocytes of patients, carriers, and healthy control subjects. We detected non-efficient actin polymerization in monocytes and neutrophils of homozygous patients compared to carriers or the healthy subjects. In monocytes, the increase in median fluorescence intensities was significantly lower in patients compared to carriers (104 vs. 213%; p < 0.01) and healthy controls (104 vs. 289%; p < 0.01). Similarly, the increase in median fluorescence intensities in neutrophils was significantly increased in the group with carriers (208%; p < 0.01) and healthy controls (238%; p < 0.01) and significantly decreased in the patient's group (94%). Our functional fMLP/phalloidin test can therefore be used as a practical tool to separate symptomatic patients from asymptomatic mutation associated to actin polymerization.

MORPHOLOGICAL AND DEVELOPMENTAL STUDY OF HUMAN FETAL THYMUS GLAND IN KUMAON REGION

Introduction: A cross sectional study was conducted to observe morphological and developmental changes occurring in thymus gland at various gestational age among human fetuses. The morphometric measurements of thymus is useful in calculating size as intrauterine growth retardation (IUGR) and cellular immune deficiency of infant is associated with a significant decrease in fetal thymic size. Materials &amp; Methods: This study was carried on medically aborted and stillborn fetuses (n=31; F=21, M=10), which were obtained from Dr. Sushila Tiwari Memorial Hospital, Haldwani, (Uttarakhand,) with due consent of parents. Fetuses were preserved in 10% formalin. Thymic gland was dissected out to observe its weight, length and thickness. Results: Thymus glands were located in superior mediastinum. On gross examination, they were of greyish pink to greyish brown in colour. Most of the glands were bilobed and few were irregular in shape. There was progressive increase in all morphometric dimensions of thymus in relation to gestational age. Growth of right lobe of thymus was comparatively more than of left lobe. Conclusion: Thymic morphometric parameters in relation to gestational age can be used as reference values in imaging studies in the prenatal and in initial perinatal stages, so we could compare the normal morphology with that of in IUGR and immune deficiency.

Rapid generation of multivirus-specific T lymphocytes for the prevention and treatment of respiratory viral infections.

Respiratory tract infections due to community-acquired respiratory viruses including respiratory syncytial virus, Influenza, parainfluenza virus 3 and human metapneumovirus are detected in up to 40% of allogeneic hematopoietic stem cell transplant recipients in whom they cause severe symptoms including pneumonia and bronchiolitis and can be fatal. Given the lack of effective antivirals and the data from our group demonstrating that adoptively transferred ex vivo-expanded virus-specific T cells can be clinically beneficial for the treatment of both latent (Epstein-Barr virus, cytomegalovirus, BK virus, human herpesvirus 6) and lytic (adenovirus) viruses, we investigated the potential for extending this immunotherapeutic approach to respiratory viruses. We now describe a system that rapidly generates a single preparation of polyclonal (CD4+ and CD8+) virus-specific T cells reactive against 12 antigens derived from 4 viruses (respiratory syncytial virus, Influenza, parainfluenza virus 3 and human metapneumovirus) that commonly cause post-transplant morbidity and mortality. With a single in vitro stimulation we consistently generated Th1-polarized T cell lines that produced multiple effector cytokines and were selectively reactive against viral-expressing targets, with no evidence of 'off target' auto- or allo-reactivity. Finally, we demonstrated the clinical relevance of these virus-specific T cells by measuring responses in transplant recipients who successfully controlled active infections. These results support the clinical importance of T cell immunity in mediating protective antiviral effects against community-acquired respiratory viruses and demonstrate the feasibility of utilizing a broad-spectrum immunotherapeutic in immunocompromised patients with uncontrolled infections.

Clinical case of of lumbar-sacral myelitis HSV2-etiology in a patient with a selective deficiency of natural killers

In this article deals with a typical clinical example of a severe form of acute neuroinfectious lesion of the spinal cord HSV2‑etiology in a patient with minor cellular immunodeficiency, who suffered from often recurrent genital herpes without adequate prophylactic antiviral treatment for the prevention of CNS damage. The patient, M., 54 years old, had acute lumbosacral myelitis, caused by herpes virus type 2, with a picture of the lower deep paraparesis, paragypesthesia, impaired pelvic organs functions. The diagnosis is confirmed on the basis of a PCR of the cerebrospinal fluid. Immunological examination revealed a selective deficiency of natural killers (CD3–CD1+CD56+ lymphocytes) — cells that are important in controlling the latent virus in the sacral sensory ganglia. A genetic survey identified a deficiency of the folate cycle that caused the minor cellular immune deficiency (MTHFR 677 С > Т homo, MTHFR 1298 А > С hetero, MTRR 66 A > G hetero). Therefore, the patient received dual treatment — acyclic analogues of guanosine to suppress viral reproduction and interferone therapy to compensate cellular immunodeficiency. This case demonstrates the feasibility of close interaction between neurologists and clinical immunologists in immunocompromised patients with neuroinfectional lesions caused by opportunistic pathogens.

Altered Gut Microbiota in HIV Infection: Future Perspective of Fecal Microbiota Transplantation Therapy.

HIV infection progressively destroys CD4+ mononuclear cells, leading to profound cellular immune deficiency that manifests as life-threatening opportunistic infections and malignancies (i.e., AIDS). Gut microbiota plays key roles in the modulation of host metabolism and gene expression, maintenance of epithelial integrity, and mediation of inflammatory and immunity. Hence, the normal intestinal microbiota plays a major role in the maintenance of health and disease prevention. In fact, a large number of studies have shown that the alteration of the gut microbiota contributes to the pathogenesis of several diseases, such as inflammatory bowel diseases, irritable bowel syndrome, metabolic diseases, anorexia nervosa, autoimmune diseases, multiple sclerosis, cancer, neuropsychiatric disorders, and cardiovascular diseases. Recently, accumulating evidence has shed light on the association of dysbiosis of gut microbiota with HIV infection. Hence, the modification of gut microbiota may be a potential therapeutic tool. Fecal microbiota transplantation may improve the conditions of patients with HIV infection by manipulating the human intestinal bacteria. However, the relevant research is very limited, and a large amount of scientific research work needs to be done in the near future.