Low Levels Of CD4 Research Articles

Characterization of Conventional Dendritic Cells and Macrophages in the Spleen Using the CSF1R-Reporter Transgenic Chickens

The spleen is a major site for the immunological responses to blood-borne antigens that is coordinated by cells of the mononuclear phagocyte system (MPS). The chicken spleen is populated with a number of different macrophages while the presence of conventional dendritic cells (cDC) has been described. However, a detailed characterization of the phenotype and function of different macrophage subsets and cDC in the chicken spleen is limited. Using theCSF1R-reporter transgenic chickens (CSF1R-tg), in which cells of the MPS express a transgene under the control elements of the chickenCSF1R, we carried out an in-depth characterization of these cells in the spleen. Immunohistological analysis demonstrated differential expression of MRC1L-B by periarteriolar lymphoid sheaths (PALS)-associatedCSF1R-tg+cells. In the chicken's equivalent of the mammalian marginal zone, the peri-ellipsoid white-pulp (PWP), we identified high expression of putative CD11c by ellipsoid-associated cells compared to ellipsoid-associated macrophages. In addition, we identified a novel ellipsoid macrophage subset that expressed MHCII, CD11c, MRC1L-B, and CSF1R but not theCSF1R-tg. In flow cytometric analysis, diverse expression of theCSF1R-tg and MHCII was observed leading to the categorization ofCSF1R-tg cells intoCSF1R-tgdimMHCIIinter−hi,CSF1R-tghiMHCIIhi, andCSF1R-tghiMHCIIintersubpopulations. Low levels of CD80, CD40, MHCI, CD44, and Ch74.2 were expressed by theCSF1R-tghiMHCIIintercells. Functionally,in vivofluorescent bead uptake was significantly higher in theCSF1R-tghiMHCIIhiMRC1L-B+cells compared to theCSF1R-tgdimandCSF1R-tghiMHCIIinterMRC1L-B+subpopulations while LPS enhanced phagocytosis by theCSF1R-tghiMHCIIintersubpopulation. The analysis of bead localization in the spleen suggests the presence of ellipsoid-associated macrophage subsets. In addition, we demonstrated the functionality ofex vivoderivedCSF1R-tg+MRC1L-BnegcDC. Finally, RNA-seq analysis of theCSF1R-tg subpopulations demonstrated that separating theCSF1R-tghisubpopulation into CD11chiand CD11cdimcells enriched for cDC and macrophage lineages, respectively, while theCSF1R-tghiMHCIIintersubpopulation was enriched for red pulp macrophages. However, our analysis could not define the cell lineage of the heterogeneousCSF1R-tgdimsubpopulation. This detailed overview of the MPS in the chicken spleen will contribute to future research on their role in antigen uptake and presentation.

Reduced Trypanosoma cruzi-specific humoral response and enhanced T cell immunity after treatment interruption with benznidazole in chronic Chagas disease.

Interruption of benznidazole therapy due to the appearance of adverse effects, which is presumed to lead to treatment failure, is a major drawback in the treatment of chronic Chagas disease. Trypanosoma cruzi-specific humoral and T cell responses, T cell phenotype and parasite load were measured to compare the outcome in 33 subjects with chronic Chagas disease treated with an incomplete benznidazole regimen and 58 subjects treated with the complete regimen, during a median follow-up period of 48 months. Both treatment regimens induced a reduction in the T. cruzi-specific antibody levels and similar rates of treatment failure when evaluated using quantitative PCR. Regardless of the regimen, polyfunctional CD4+ T cells increased in the subjects, with successful treatment outcome defined as a decrease of T. cruzi-specific antibodies. Regardless of the serological outcome, naive and central memory T cells increased after both regimens. A decrease in CD4+ HLA-DR+ T cells was associated with successful treatment in both regimens. The cytokine profiles of subjects with successful treatment showed fewer inflammatory mediators than those of the untreated T. cruzi-infected subjects. High levels of T cells expressing IL-7 receptor and low levels of CD8+ T cells expressing the programmed cell death protein 1 at baseline were associated with successful treatment following benznidazole interruption. These findings challenge the notion that treatment failure is the sole potential outcome of an incomplete benznidazole regimen and support the need for further assessment of the treatment protocols for chronic Chagas disease.

Serum CD4 Is Associated with the Infiltration of CD4+T Cells in the Tumor Microenvironment of Gastric Cancer.

Serum CD4, CD8, and CD19 are markers of systemic inflammation. However, there is little evidence on the influence of inflammation on the tumor microenvironment and the prognostic indicators of gastric cancer (GC). In this study, two hundred and eight patients who underwent radical gastrectomy for GC were included. Preoperative peripheral blood samples were used to analyze Serum CD4, CD8, and CD19. The optimal cutoff levels for CD4, CD8, and CD19 were defined by receiver operating characteristic curve analysis (CD4 = 38.85%, CD8 = 14.35%, and CD19 = 7.40%). The areas with specific CD4+T cells, CD8+T cells, and CD19+B cells within the tumor microenvironment were measured in paraffin sections by immunohistochemistry and analyzed by Image-Pro Plus. 94 patients had low CD4, and 124 patients had high CD4 levels. 31 patients had low CD8, and 187 patients had high CD8 levels. 64 patients had low CD19, and 154 patients had high CD19 levels. Infiltration of CD4+T cells was associated with serum CD4 (P < 0.001). Serum CD4 and CD19 and the infiltration of CD4+T cells, CD8+T cells, and CD19+B cells were significant in predicting the prognosis of GC. Low CD4 level, infiltration of CD8+T cells, and high infiltration of CD4+T cells and CD19+B cells were correlated with worse overall survival in multivariate analysis. Collectively, our results provide evidence that serum CD4 is associated with the infiltration of CD4+T cells in the tumor microenvironment, which indicates the prognostic value of systemic inflammation in GC.

Analysis on death trend in AIDS patients and related risk factors in China

Objective: To understand the basic characteristics of death cases, analyze the death trends in AIDS patients and the risk factors in China and provide evidence for the development of AIDS prevention and control strategy. Methods: The data were collected from the national basic information system of HIV/AIDS. The information of the cases in AIDS phase were used. The death number and mortality trends in AIDS cases were described, and Cox Proportion Hazards Regression Model was constructed to assess hazard ratios (HR) for independent variables. Results: By the end of 2019, a total of 582 472 AIDS cases, including 168 391 deaths, had been reported in China. Among the death cases, males accounted for 76.8% (129 343/168 391), heterosexual contact was the main transmission route, accounting for 60.9% (102 516/168 391). The proportion of the death cases who had ever received ART was 54.0% (90 888/168 391). The inter-quartile (P25, P75) of first CD4＋T cells counts (CD4) was 34 cells/μl, 240 cells/μl. Up to 43.5% (73 191/168 391) of the deaths occurred within one year after diagnosis. From 2007 to 2019, the annual death number increased from 5 485 to 18 737, the mortality rates decreased form 10.9%% to 4.3%. The average time interval from diagnosis to death ranged from 1.4 year to 4.0 years, showing increase trend by year. The results of Cox regression analysis showed that older age (50- years old: HR=1.50; ≥65 years old: HR=2.00), being male (HR=1.44)、being in minority ethnic group (HR=1.10), having lower first CD4 levels (0- cells/μl, HR=2.73;200- cells/μl, HR=1.33; 350- cells/μl,HR=1.13), heterosexual transmission route (HR=1.64) and injecting drug use (HR=1.79) were the risk factors related to deaths in AIDS patients. The higher educational levels (junior middle school: HR=0.86, senior high school and above: HR=0.59) and receiving antiviral treatrment (HR=0.09) were protective factors. Conclusions: The number of death cases increased, meanwhile the mortality rates decrease year by year in AIDS patients in China during 2007-2019. It is necessary to strengthen the early detection and treatment of AIDS to reduce the mortality.

Immune Reconstitution Inflammatory Syndrome and Hodgkin’s Lymphoma

Immune reconstitution inflammatory syndrome (IRIS) is defined as a clinically significant exacerbation of known oligosymptomatic serious, more often infectious, diseases with considerably increased CD4+ T-lymphocyte count in response to highly active anti-retroviral therapy (HAART) of HIV infection. The review comprehensively discusses tuberculosis issues in HIV-positive HAART recipients. Related recommendations contain strict guidelines on compulsory treatment of tuberculosis prior to HAART assignment. Similar recommendations for specific therapy preceding HAART are provided for other opportunistic infections (mycotic and cryptococcal infections, parasitosis, molluscum contagiosum, toxoplasmosis, herpes-zoster virus, leishmaniasis, syphilis, and lepra). Without prior specific therapy of an opportunistic infection its exacerbation with pronounced symptoms and signs on HAART can be fatal for the patient. Lymphomas including Hodgkin’s lymphoma (HL) are dealt with in the context of the same challenge. However, what remains unclear is the specificity of targeted T-lymphocytes in the microenvironment to hitherto unclarified cause-specific antigens of the tumor. As opposed to other malignant lymphoid tumors arising with low level of CD4+ T-lymphocytes, HL develops when the level of CD4+ T-lymphocytes is increased in response to HAART in HIV-positive patients during the first months of anti-retroviral therapy. HL is diagnosed in 8 % of HIV-positive off-HAART subjects. After HAART assignment the HL incidence goes up to 17 %. Therefore, IRIS can be considered the main challenge in the study of etiology and pathogenesis of HL in HIV-positive patients. In this context, the demand to extend the research in this field becomes not only obvious but crucial for practical applications.

Prevalence of Opportunistic and other Intestinal Parasites Infections and its Associated Factors among HIV/AIDS Patients attending at Dessie Comprehensive Specialized Hospital, Northeast Ethiopia

Introduction: Human Immuno Deficiency Virus (HIV) deplete CD4+ T cell in human and weaken the immune system, this makes HIV positive patients more susceptible to parasitic and other opportunistic infection. Intestinal parasitic infection plays vital role in the prognosis of people living with HIV/AIDS. This study was aimed to determine the magnitude of intestinal parasitic infection and its determinant among people living HIV/AIDS. Method: A cross-sectional study was conducted from February 2020 to April 2020. A total of 223 study participants were recruited using simple random sampling. A Pre-tested questionnaire was used to collect socio-demographic and other risk factor data. A stool sample was collected to detect parasitic infection using wet mount, formol-ether concentration and Modified ziehl-neelsen technique. The whole blood sample was collected to determine CD4+ T cell count using BD FACSCount™ System. Data was entered into Epi Data version 3.1 and analyzed using SPSS version 20. Results: Out of 223 participants 166 (74.4%) were males. The mean age of the study participants was 37.9 years old with the majority being found in the age group 26-49 years (64.1%). The overall prevalence of intestinal parasitic infection was found to be 38.1%. The prevalence was significantly higher in males (23.7%) than in females (p≤0.002). The most predominant parasite detected was G. lamblia (40%) followed by E. histolytica (32.9%). The prevalence of opportunistic infection was 2.24%. The detected opportunistic parasites were S. stercoloaris, Cryptosporidium spps and I. beli. Sex, residence and low CD4+ T cell counts were significantly associated with the prevalence intestinal parasitic infection among HIV/ AIDS patients. Conclusion: The finding showed intestinal parasitic infections being a major health problem in HIV patients. Low-level CD4 T cell is a risk factor for the high prevalence of parasitic infection. The high prevalence of intestinal parasitic infection indicates the need of routine investigation of the infection that will aid for rapid therapeutic management.

509. Comparision of CD4+ T Cells in Patients with Severe vs Critical COVID -19

BackgroundOver the past few years, it has been shown that T cells play an essential role in antiviral immunity, in the course of the COVID-19 pandemic some studies reported an association between lymphocytopenia and exhaustion of the surviving remaining T cells which are apparently functional in patients with acute COVID-19, specially in those with severe forms of presentation. Some studies have reported an association where less than 800 CD4 + T cells are negatively related to the survival of seriously ill patients with COVID -19.MethodsWe included 19 patients admitted to our hospital (ABC Medical Center) from May 7 to 15, 2020 with a confirmed diagnosis of COVID-19 and were randomized into 2 groups according to the severity of the presentation (severe or critical) A determination of CD4 + T cells was made at admission, we also reported the need for invasive mechanical ventilation at some point of the hospitalization for each group, all patients were followed until their hospital discharge. One patient was excluded because he was still admitted at the time of the analysis.ResultsOf the 18 patients included, 9 (50%) fulfilled criteria of severe and 9 (50%) of critical. The mean of CD4 + T cell was 455 (256–697) for the severe and 285.44 (145–430) for the critical (CI 95% P 0.46), the determination of CD8+ T cell was 212 (88–392) for the severe and 201 (59–534) for the critical (CI 95% P 1.19), of the critical patients 8 (88.9%) required invasive mechanical ventilation and only one non-invasive mechanical ventilation, while the severe patients only required support with supplemental oxygen by nasal cannula (9 (100%)).The mean lenght of hospitalization was 12.73 days (3–34) and all the patients survived until they were discharged home.ConclusionAs it has been reported in some studies, the pathogenesis of SARS-CoV-2 infection in humans is associated with a reduction and functional exhaustion of T cells in patients with COVID-19.In this study we presume that lower levels of CD4+T cells can be associated with critical forms of COVID 19 as the majority of critical patients in our report had < 300 CD4 +T cell count, while we need further studies with a greater number of patients and follow-up to establish reliable determinations, we propose than the levels of CD4+T cell count could be use as a good predictor of severity in COVID-19Disclosures All Authors: No reported disclosures

Activation of T helper cells in sentinel node predicts poor prognosis in oral squamous cell carcinoma

Recurrence in oral squamous cell carcinoma (OSCC) significantly reduces overall survival. Improved understanding of the host’s immune status in head and neck cancer may facilitate identification of patients at higher risk of recurrence and improve patients’ selection for ongoing clinical trials assessing the effectiveness of immune checkpoint inhibitors (CPI). We aimed to investigate Sentinel Node-derived T-cells and their impact on survival. We enrolled prospectively 28 OSCC patients treated at Karolinska University Hospital, Stockholm, Sweden with primary tumour excision and elective neck dissection. On top of the standard treatment, the enrolled patients underwent sentinel node procedure. T cells derived from Sentinel nodes, non-sentinel nodes, primary tumour and PBMC were analyzed in flow cytometry. Patients who developed recurrence proved to have significantly lower level of CD4+ CD69+ in their sentinel node (31.38 ± 6.019% vs. 43.44 ± 15.33%, p = 0.0103) and significantly higher level of CD8+ CD HLA-DR+ (38.95 ± 9.479% vs. 24.58 ± 11.36%, p = 0.0116) compared to disease-free individuals. Survival analysis of studied population revealed that patients with low proportion of CD4+ CD69+ had significantly decreased disease-free survival (DFS) of 19.7 months (95% CI 12.6–26.9) compared with 42.6 months (95% CI 40.1–45.1) in those with high CD4+ CD69+ proportion in their Sentinel Nodes (log-rank test, p = 0.033). Our findings demonstrate that characterization of T-cell activation in Sentinel Node serves as a complementary prognostic marker. Flow cytometry of Sentinel Node may be useful in both patients’ surveillance and selection for ongoing CPI clinical trials in head and neck cancer.

Hyperlipasemia and Potential Pancreatic Injury Patterns in COVID-19: A Marker of Severity or Innocent Bystander?

Hyperlipasemia and Potential Pancreatic Injury Patterns in COVID-19: A Marker of Severity or Innocent Bystander?

B7-H4 Inhibits the Development of Primary Sjögren's Syndrome by Regulating Treg Differentiation in NOD/Ltj Mice.

Background This study is aimed at exploring the role of B7-H4 in the pathogenesis of primary Sjögren's syndrome (pSS) in NOD/Ltj mouse. Methods B7-H4 expression in salivary glands was examined by IHC, and lymphocyte infiltration was showed by H&E. Next, anti-B7-H4 mAb or immunoglobulin isotype was injected into NOD/Ltj mice. Cytokine levels were measured by quantitative RT-PCR, and immunoglobulins were measured by ELISA. T cell subsets were analyzed by flow cytometry. Last, we treated NOD/Ltj mice with B7-H4Ig and control Ig. CD4+Foxp3+ T cells were assessed by immunohistochemistry. Two-tailed Student's t-tests were used to detect the statistical difference in various measures between the two groups. Results B7-H4 expression was remarkably reduced in salivary glands of NOD/Ltj mice at 15 weeks compared with the NOD/Ltj mice at 8 weeks. Anti-B7-H4 mAb treatment increased lymphocyte infiltration in salivary glands. Inflammatory cytokines including IL-12, IL-18, IL-1α, TNF-α, IFN-α, and BAFF were upregulated markedly in anti-B7-H4 mAb-treated mice compared to IgG isotype-treated mice. Flow cytometry analysis showed that anti-B7-H4 mAb-treated mice had lower levels of CD4+Foxp3+/CD4+ T cells in spleen. Moreover, Foxp3 mRNA levels of salivary glands were diminished in anti-B7-H4 mAb-treated mice. Flow cytometry analysis showed that anti-B7-H4 mAb inhibited CD4+Foxp3+/CD4+ T cell production, while B7-H4Ig would promote naïve CD4+ T into Treg differentiation. Administration with B7-H4Ig displayed significantly decreased lymphocyte infiltration in salivary glands and low levels of total IgM and IgG in serum. Analysis of inflammatory cytokines in salivary glands after B7-H4Ig treatment revealed that the mRNA levels of IL-12, IL-6, IL-18, IL-1α, TNF-α, and IFN-α were significantly downregulated in B7-H4Ig-treated mice compared to control Ig treatment. B7-H4Ig-treated mice had significantly higher levels of CD4+Foxp3+/CD4+ T cells in spleen. IHC in salivary gland revealed that CD4+Foxp3+ T cells of B7-H4Ig treatment mouse were more than control Ig treatment. Conclusions Our findings implicate that B7-H4 has a protective role for salivary gland epithelial cells (SGECs) and therapeutic potential in the treatment of pSS.

Causes of fever in Tanzanian adults attending outpatient clinics: a prospective cohort study

ObjectivesExploring fever aetiologies improves patient management. Most febrile adults are outpatients, but all previous studies were conducted in inpatients. This study describes the spectrum of diseases in adults attending outpatient clinics in urban Tanzania. MethodsWe recruited consecutive adults with temperature ≥38°C in a prospective cohort study. We collected medical history and performed a clinical examination. We performed 27 364 microbiological diagnostic tests (rapid tests, serologies, cultures and molecular analyses) for a large range of pathogens on blood and nasopharyngeal samples. We based our diagnosis on predefined clinical and microbiological criteria. ResultsOf 519 individuals, 469 (89%) had a clinically or microbiologically documented infection and 128 (25%) were human immunodeficiency virus (HIV) -infected. We identified 643 diagnoses: 264 (41%) acute respiratory infections (36 (5.6%) pneumonia, 39 (6.1%) tuberculosis), 71 (11%) infections with another focus (31 (4.8%) gastrointestinal, 26 (4.0%) urogenital, 8 (1.2%) central nervous system) and 252 (39%) infections without focus (134 (21%) dengue, 30 (4.7%) malaria, 28 (4.4%) typhoid). Of the 519 individuals, 318 (61%), 179 (34%), 30 (6%) and 15 (3%), respectively, had a viral, bacterial, parasitic and fungal acute infection. HIV-infected individuals had more bacterial infections than HIV-negative (80/122 (66%) versus 100/391 (26%); p < 0.001). Patients with advanced HIV disease had a higher proportion of bacterial infections (55/76 (72%) if CD4 ≤200 cells/mm3 and 25/52 (48%) if CD4 >200 cells/mm3, p 0.02). ConclusionsViral diseases caused most febrile episodes in adults attending outpatient clinics except in HIV-infected patients. HIV status and a low CD4 level strongly determined the need for antibiotics. Systematic HIV screening is essential to appropriately manage febrile patients.

IL-27 Regulated CD4+IL-10+ T Cells in Experimental Sjögren Syndrome.

Interleukin 27 (IL-27) plays diverse immune regulatory roles in autoimmune disorders and promotes the generation of IL-10–producing CD4+ T cells characterized by producing the immunosuppressive cytokine IL-10. However, whether IL-27 participates in pathological progress of Sjögren syndrome (SS) through regulating CD4+IL-10+ T cells remains unknown. Here we aimed to explore the potential role of IL-27 and CD4+IL-10+ T cells in the pathogenesis of SS. The IL-27 gene knockout non-obese diabetic (Il-27−/−NOD) mice were generated and injected with exogenous IL-27. Exogenous injection of IL-27 and neutralization of IL-27 with anti–IL-27 antibody in NOD mice were performed. The histopathologic changes in submandibular glands, lacrimal glands and lung, salivary flow rate, and percentages of CD4+IL-10+ T cells were determined. And, ovalbumin-immunized C57L/B6 mice were injected with IL-27 to detect the percentage of CD4+IL-10+ T cells. In vitro, splenic naive T cells from C57L/B6 mice were cultured with IL-27 for 4 days to induce the differentiation of CD4+IL-10+ T cells. In addition, IL-27, IL-10, and CD4+IL-10+ T cells were determined in health control and SS patients. The results showed that Il-27−/−NOD mice had more severe disease and lower level of CD4+IL-10+ T cells than control mice. And IL-27 promoted the generation and differentiation of CD4+IL-10+ T cells in vivo and in vitro significantly. In agreement with the findings in the SS-like mice, patients with SS showed lower levels of IL-27, IL-10, and CD4+IL-10+ T cells. Our findings indicated that IL-27 deficiency aggravated SS by regulating CD4+IL-10+ T cells. Targeting IL-27 and CD4+IL-10+ T cells may be a novel therapy for patients with SS.

Expression and clinical significance of LAG-3, FGL1, PD-L1 and CD8+T cells in hepatocellular carcinoma using multiplex quantitative analysis

BackgroundFibrinogen-like protein 1 (FGL1)—Lymphocyte activating gene 3 (LAG-3) pathway is a promising immunotherapeutic target and has synergistic effect with programmed death 1 (PD-1)/programmed death ligand 1 (PD-L1). However, the prognostic significance of FGL1-LAG-3 pathway and the correlation with PD-L1 in hepatocellular carcinoma (HCC) remain unknown.MethodsThe levels of LAG-3, FGL1, PD-L1 and cytotoxic T (CD8+T) cells in 143 HCC patients were assessed by multiplex immunofluorescence. Associations between the marker’s expression and clinical significances were studied.ResultsWe found FGL1 and LAG-3 densities were elevated while PD-L1 and CD8 were decreased in HCC tissues compared to adjacent normal liver tissues. High levels of FGL1 were strongly associated with high densities of LAG-3+cells but not PD-L1. CD8+ T cells densities had positive correlation with PD-L1 levels and negative association with FGL1 expression. Elevated densities of LAG-3+cells and low levels of CD8+ T cells were correlated with poor disease outcome. Moreover, LAG-3+cells deteriorated patient stratification based on the abundance of CD8+ T cells. Patients with positive PD-L1 expression on tumor cells (PD-L1 TC+) tended to have an improved survival than that with negative PD-L1 expression on tumor cells (PD-L1 TC−). Furthermore, PD-L1 TC− in combination with high densities of LAG-3+cells showed the worst prognosis, andPD-L1 TC+ patients with low densities of LAG-3+cells had the best prognosis.ConclusionsLAG-3, FGL1, PD-L1 and CD8 have distinct tissue distribution and relationships with each other. High levels of LAG-3+cells and CD8+ T cells represent unfavorable and favorable prognostic biomarkers for HCC respectively.

Prenatal Management of Congenital Human Cytomegalovirus Infection in Seropositive Pregnant Patients Treated with Azathioprine

Human cytomegalovirus (HCMV) is the leading infectious agent causing congenital disabilities. The risk of HCMV transmission to the fetus in pregnant women receiving immunosuppressive agents is unknown. We describe two cases of pregnant women with evidence of pre-conception HCMV protective immunity receiving azathioprine for ulcerative colitis or systemic lupus erythematosus. Both women reactivated the HCMV and transmitted the infection to the fetuses. One newborn showed unilateral hearing deficits and brain abnormalities while the other was asymptomatic. The mother of the symptomatic newborn had low levels of total and HCMV-specific blood CD4+ T cells. Women receiving immunosuppressive agents deserve information about the risk of HCMV congenital infection and should be monitored for HCMV infection during pregnancy. Their newborns should be screened for HCMV congenital infection.

Immune status, and not HIV infection or exposure, drives the development of the oral microbiota

Even with antiretroviral therapy, children born to HIV-infected (HI) mothers are at a higher risk of early-life infections and morbidities including dental disease. The increased risk of dental caries in HI children suggest immune-mediated changes in oral bacterial communities, however, the impact of perinatal HIV exposure on the oral microbiota remains unclear. We hypothesized that the oral microbiota of HI and perinatally HIV-exposed-but-uninfected (HEU) children will significantly differ from HIV-unexposed-and-uninfected (HUU) children. Saliva samples from 286 child-participants in Nigeria, aged ≤ 6 years, were analyzed using 16S rRNA gene sequencing. Perinatal HIV infection was significantly associated with community composition (HI vs. HUU—p = 0.04; HEU vs. HUU—p = 0.11) however, immune status had stronger impacts on bacterial profiles (p < 0.001). We observed age-stratified associations of perinatal HIV exposure on community composition, with HEU children differing from HUU children in early life but HEU children becoming more similar to HUU children with age. Our findings suggest that, regardless of age, HIV infection or exposure, low CD4 levels persistently alter the oral microbiota during this critical developmental period. Data also indicates that, while HIV infection clearly shapes the developing infant oral microbiome, the effect of perinatal exposure (without infection) appears transient.

Therapeutic effects of conditioned – DPBS from amniotic stem cells on lactating cow mastitis

ObjectiveBovine mastitis results in economic loss due to decrease in milk production. Antibiotic ointments are commonly used for treating. However, residue and anti-microbial resistance warranted attention progressively. Fortunately, stem cell anti-inflammatory properties and paracrine expression of cytokines accelerates wound healing and suppresses inflammatory reactions in mastitis. The objective of this study is to use the conditioned-Dulbecco's pluripotent stem cells (DPBS) from amniotic membrane stem cells (AMSCs) in treating bovine mastitis. Materials and methodsThe cows with mastitis were divided into two groups. In antibiotic control group, the cows were given tetraneomycin ointment. In conditioned-DPBS of AMSCs treatment group, amniotic membrane was collected for AMSCs after delivery. With expression of surface antigen and potential of tri-linage differentiation, AMSCs were injected into mammary glands. Then, milk was sampled every three days to monitor the effect of both treatments. The quality of milk was measured with pH, titratable acidity, free calcium ions and somatic cell count. ResultsOur results demonstrated the Bovine AMSCs expressed CD44, low levels of CD4 and no CD105. Bovine AMSCs demonstrated the differentiation capability in the tri-cell lineages. Mastitis treatment with conditioned-DPBS from AMSCs (experimental group) and conventional antibiotics (control group) showed insignificant difference in pH value and titratable acidity. The level of ionic calcium concentration in the conditioned-DPBS group decreased from 3rd day to 12th day, while the level in the antibiotic group decreased from 0 day to 12th day. The somatic cell number was similar in both groups, which meet the standard of Taiwan milk collection. ConclusionIn conclusion, conditioned-DPBS from bovine AMSCs has the therapeutic potential to treat bovine mastitis and may replace antibiotics therapy in the future.

Justification of the integrated approach to management of hiv-infected patients with comorbid pathology of the digestive system

Introduction. Among the main strategic and operational goals of the State Strategy for Combating HIV / AIDS, Tuberculosis and Viral Hepatitis by 2030 in Ukraine is to ensure comprehensive access to HIV treatment, increase the effectiveness of monitoring and support of treatment of both opportunistic infections and other somatic conditions in HIV-infected patients. The key role of family physicians in the detection and treatment of many chronic gastrointestinal complications in HIV-infected patients is recognized.&#x0D; Purpose of the study. To increase the efficiency of early diagnosis and tactics of integrated management of HIV-infected patients of family physicians (FP) by clarifying the peculiarities of HIV infection in the presence of comorbid pathology of the digestive system (DS) and creating an algorithm for providing medical care to these patients.&#x0D; Material and methods. The research was conducted on the basis of five regional HIV / AIDS centers of Ukraine during 2017-2019. Randomly selected 342 adult HIV-infected patients were divided into two groups - with concomitant lesions and without concomitant gastrointestinal lesions. The following research methods were used: general clinical and laboratory biochemical, molecular genetic, immunological, enzyme-linked immunosorbent, instrumental (FGDS, chest radiography, abdominal ultrasound, computed tomography of the chest and / or abdominal cavity), analysis of primary medical records, consultations related specialists according to the indications, questionnaires, statistical methods.&#x0D; Results and discussion. It was found that the incidence of gastrointestinal pathology in patients with II, III and IV clinical stages of HIV infection was significantly higher than in patients with stage I, significantly more often associated with tuberculosis, candidiasis, kidney disease and HIV encephalopathy and was combined. Manifestations of asthenovegetative and dyspeptic syndromes, weight loss, anemia and leukopenia, increased activity of liver enzymes, low levels of CD4 + lymphocytes and preservation of viral load on antiretroviral therapy were significantly more common in HIV-infected patients with gastrointestinal pathology. In the presence of pathology of the digestive system, replacements, breaks and side effects of antiretroviral therapy were significantly more frequent. The results of physicians survey analysis showed the role of family physicians in the current examination and management of HIV-infected people with comorbid pathology of the digestive system.&#x0D; Conclusions. To detect diseases of the organs of the DS in HIV-infected people, it is necessary to conduct a comprehensive laboratory and instrumental examination, taking into account the possibility of combined pathology. The proposed algorithm of integrated management of HIV-infected patients with comorbid pathology of DS by FPs, taking into account the most informative clinical and laboratory criteria, allows to increase the effectiveness of early diagnosis and tactics of integrated management of HIV-infected by FPs.

Interferon lambda rs368234815 \u0394G/\u0394G is associated with higher CD4+:CD8+ T-cell ratio in treated HIV-1 infection

BackgroundThe objectives of this study were to investigate the relationships between polymorphisms at the interferon lambda (IFNL) locus and CD4+:CD8+ ratio normalisation in people living with HIV (PLWH) on effective antiretroviral therapy (ART); and to examine whether these polymorphisms influence the composition of T lymphocyte compartments in long-term treated HIV-1 infection.MethodsA cross-sectional study in PLWH enrolled into the Mater Immunology study. We performed IFNL genotyping on stored samples and evaluated the association of IFNL single-nucleotide polymorphisms (rs368234815 and rs12979860) with CD4+:CD8+ ratio normalization (> 1) and expanded CD4+ and CD8+ T-cell subsets; CD45RO+CD62L+ (central-memory), CD45RO+ CD62L−(effector-memory) and CD45RO−CD62L+ (naïve), using logistic and linear regression models, respectively.Results190 ambulatory PLWH recruited to the main study, 143 were included in the analysis (38 had no stored DNA and 9 no T-lymphocyte subpopulation). Of 143 included, the median age (IQR) was 45(39–48) years, 64% were male and 66% were of Caucasian ethnicity. Heterosexual-contact (36%), injecting drug-use (33%) and men who have sex with men (24%) were the most presented HIV-transmission risk groups. The majority of subjects (90.2%) were on ART with 79% of the cohort having an undetectable HIV-RNA (< 40 copies/ml) and the time since ART initiation was 7.5 (3.7–10.4) year. rs368234815 and rs12979860 displayed similar allelic frequencies, with minor alleles ΔG and T representing 39% and 42%, respectively, of circulating alleles. rs368234815 ΔG/ΔG minor homozygotes were significantly associated with increased odds for attaining a normalised CD4+:CD8+ ratio compared to rs368234815 T/T major homozygotes in PLWH virologically suppressed on effective ART (OR = 3.11; 95% CI [1.01:9.56]). rs368234815 ΔG/ΔG homozygosity was also significantly associated with lower levels of CD4+ effector memory T-cells (regression coefficient: − 7.1%, p = 0.04) and CD8+ naïve T-cell subsets were significantly higher in HIV-1 mono-infected PLWH with rs368234815 ΔG/ΔG (regression coefficient: + 7.2%, p = 0.04).ConclusionsIn virally-suppressed, long-term ART-treated PLWH, rs368234815 ΔG/ΔG homozygotes were more likely to have attained normalisation of their CD4+:CD8+ ratio, displayed lower CD4+ effector memory and higher naive CD8+ T-cells. Further studies are needed to replicate our findings in other, larger and more diverse cohorts and to determine the impact of IFNL genetic-variation on CD4+:CD8+ ratio normalisation and clinical outcomes in PLWH.

A protocol for rapid monocyte isolation and generation of singular human monocyte-derived dendritic cells.

The monocyte-derived dendritic cells (moDCs) are a subset of dendritic cells widely used in immunological studies as a convenient and easy approach after isolation of mononuclear cells directly from peripheral blood mononuclear cells (PBMC). Both the purification and cell culture of monocytes impact on the differentiation of monocytes into moDCs. The methodology to isolate and differentiate monocytes into moDCs is still controversial. We aimed to compare three different protocols for monocyte isolation from PBMC: 1) Cold-aggregation; 2) Percoll gradient; and 3) Magnetic beads cell-enrichment. Additionally we also compared four different monocyte differentiation and culture techniques: 1) Cell culture media; 2) Serum sources; 3) required GM-CSF and IL-4 concentrations; 4) Cell culture systems. We used flow cytometry analysis of light scattering and/or expression of pan surface markers, such as CD3, CD14 and CD209 to determine isolation/differentiation degree. Purified PBMC followed by two steps of cold aggregation, yielded cell viability around 95% with poor monocyte enrichment (monocytes increase vs. lymphocytes reduction was not statistically significant, p>0.05). Conversely, monocyte isolation from PBMC with discontinuous Percoll gradient generated around 50% cell viability. Albeit, we observed a significant reduction (p≤0.05) of lymphocytes contaminants. The magnetic beads cell-enrichment yield cell viability higher than 95%, as high as a significant lymphocyte depletion (p≤0.005) when compared to all other techniques employed. The moDCs showed better differentiation based on increased CD209 expression, but lower CD14 levels, when cells were cultured in RPMI medium plus 500IU/mL of both GM-CSF and IL-4 in a semi-adherent fashion. Serum sources showed no influence on the culture performance. In conclusion, the magnetic beads cell-enrichment showed superior cell viability, indicating that this approach is a better choice to isolate monocytes, and moDCs cultured afterwards in appropriate medium, serum, cytokines and culture system might influence the monocytes differentiation into moDC.

Pancreatic Injury Patterns in Patients With Coronavirus Disease 19 Pneumonia

Pancreatic Injury Patterns in Patients With Coronavirus Disease 19 Pneumonia