Enumeration Of CD4 Research Articles

Accurate and reproducible enumeration of CD4 T cell counts and Hemoglobin levels using a point of care system: Comparison with conventional laboratory based testing systems in a clinical reference laboratory in Cameroon.

Measurements of CD4 T cells and hemoglobin (Hb) are conventionally used to determine the immunological state and disease progression for HIV-infected patients. We obtained a small lightweight point-of-care device, the BD FACSPrestoTM in order to demonstrate its ability to deliver CD4 and Hb analysis in comparison with two larger clinical machines the BDFACSCantoTM analyzer and Sysmex XN 1000 haematology analyzer. The advantages of using the POC device include access to HIV patient data in remote and in resource limited settings. The analytical performance of the BD FACSPrestoTM, compared with the FACSCantoTM II flow cytometer and the Sysmex XN 1000 haematology analyzer was evaluated by testing 241 routine clinical specimens collected in EDTA tubes from patients attending the Immunology and Microbiology laboratory of Chantal BIYA International Reference Centre (Yaounde, Cameroon) between January and May 2016. The mean in absolute counts and percentage of CD4 T cells was 606 cells/mL and 25% respectively via the FACSPrestoTM, and 574 cells/mL and 24% respectively via the BD FACSCantoTM II. The mean concentration of Hb levels was 11.90 on the Sysmex XN 1000 and 11.45 via the BD FACSPrestoTM, A high correlation (R2 = 0.95, P < 0.001) of Hb level measurements was noted between the BD FACSPrestoTM and Sysmex XN 1000 hematology analyzer. Overall, a Bland-Altman plot of the differences between the two methods showed an excellent agreement for absolute and percentage CD4 counts and hemoglobin measurements between POC and conventional methods evaluated here. Furthermore, the study demonstrated the ease of use of the BD FACSPrestoTM POC technology in remote areas. The BD FACPrestoTM is a suitable tool for CD4 enumeration in resource-limited settings, specifically providing a deployable, reliable POC testing option. The BD FACSPrestoTM performed appropriately in comparison to the conventional reference standard technologies. The BD FACSPrestoTM, system provides accurate, reliable, precise CD4/%CD4/Hb results on venous blood sampling. The data showed good agreement between the BD FACSPrestoTM, BD FACSCantoTM II and Sysmex XN 1000 XN 1000 systems.

Lipid Profile and Cd4+ - T – Lymphocytes Counts in Naïve Human Immunodeficiency Virus Infected Pre – Menopausal Females.

This study was aimed at determining the serum lipid levels in naive HIVinfected pre – menopausal females and in the progression of HIV/AIDS. In a bid to determine lipid abnormalities or otherwise in naïve HIV – infected pre-menopausal females, 80 HIV- positive females aged between twenty to thirty five years at the PEPFAR, HIV clinic, University of Nigeria Teaching Hospital(UNTH), Enugu and 80 apparently healthy non-HIV positive agematched female volunteers that served as the control group were studied. The medical Ethical Committee of the UNTH, Ituku-Ozalla, Enugu, approved the study protocol and the patients’ informed consents were obtained prior to sample collection. The blood samples were collected using standard methods. HIV re-screening and confirmation were carried out using WHO HIV test algorithm. Automated method using Flow cytometry was used for the enumeration of CD4+ T Lymphocytes while lipid profile of the test subjects and the control group were carried out using standard methods. The results were expressed as mean ± standard deviation. The data were analyzed using graph pad Instat version 3.01, Graph pad software, San Diego, California, USA. The test of significance was based on probability (p) values of &lt; 0.05. Comparisons between groups were performed using One-way-ANOVA, Bonferroni’s multiple comparison test and student ttests depending on the number of variables. Serum total cholesterol, HDL and LDL levels of the patients were significantly lower (P &lt; 0.05) than the control groups and all decreased progressively with the progression of HIV/AIDS. However, triacylglylerols and VLDL levels were significantly higher (P &lt; 0.05) than the controls and all increased with the progression of HIV/AIDS.

SARS-CoV-2 T-Cell Responses in Allogeneic Hematopoietic Stem Cell Recipients following Two Doses of BNT162b2 mRNA Vaccine.

Background: At variance to humoral responses, cellular immunity after anti-SARS-CoV-2 vaccines has been poorly explored in recipients of allogeneic hematopoietic stem-cell transplantation (Allo-HSCT), especially within the first post-transplant years where immunosuppression is more profound and harmful. Methods: SARS-CoV-2 Spike protein-specific T-cell responses were explored after two doses of BNT162b2 mRNA vaccine in 45 Allo-HSCT recipients with a median time from transplant of less than 2 years by using INF-γ ELISPOT assay and flow-cytometry enumeration of CD4+ and CD8+ T lymphocytes with intracellular cytokine production of IFN-γ and TNF-α. Results: A strong TNF-α+ response from SARS-CoV-2-specific CD4+ T-cells was detected in a majority of humoral responders (89%) as well as in a consistent population of non-humoral responders (40%). Conclusions: T-cells are likely to participate in protection against COVID-19 viral infection, even in the absence of detectable antibody response, especially in the first years post-transplant in Allo-HSCT recipients.

Microparticle-tagged image-based cell counting (ImmunoSpin) for CD4\u2009+\u2009T cells

Affordable point-of-care (POC) CD4 + T lymphocyte counting techniques have been developed as alternatives to flow cytometry-based instruments caring for patients with human immunodeficiency virus (HIV)-1. However, POC CD4 enumeration technologies can be inaccurate. Here, we developed a microparticle-based visual detector of CD4 + T lymphocytes (ImmunoSpin) using microparticles conjugated with anti-CD4 antibodies, independent of microfluidic or fluorescence detection systems. Visual enumeration of CD4 + T cells under conventional light microscope was accurate compared to flow cytometry. Microparticle-tagged CD4 + T cells were well-recognized under a light microscope. ImmunoSpin showed very good precision (coefficients of variation of ImmunoSpin were ≤ 10%) and high correlation with clinical-grade flow cytometry for the enumeration of CD4 + T cells (y = 0.4232 + 0.9485 × for the %CD4 + T cell count, R2 = 0.99). At thresholds of 200 and 350 cells/µL, there was no misclassification of the ImmunoSpin system compared to the reference flow cytometry. ImmunoSpin showed clear differential classification of CD4 + T lymphocytes from granulocytes and monocytes. Because non-fluorescence microparticle-tags and cytospin slides are used in ImmunoSpin, they can be applied to an automatic digital image analyzer. Slide preparation allows long-term storage, no analysis time limitations, and image transfer in remote areas.Graphical abstract

Inactivated SARS-CoV-2 vaccines elicit immunogenicity and T-cell responses in people living with HIV

BackgroundsTo date, the effects of SARS-CoV-2 vaccines on people living with HIV (PLWH) were mainly focused on messenger RNA (mRNA) and adenovirus vector-based vaccines, and little is known about the effects of inactivated virus-based vaccine. This study was designed to determine the effects of inactivated SARS-CoV-2 vaccines on PLWH. MethodsTwenty-four HIV-positive individuals and 24 healthy donors (HD) were respectively recruited from Malipo Country People's Hospital and community in Kunming city. Enumeration of lymphocyte and CD4+CD45RO+ memory T cells were evaluated by flow cytometry. Competitive ELISA was used to measure the level of Anti-SARS-CoV-2 neutralization antibody. Spearman or Pearson correlation analysis was used to analyze the relationship between laboratory indicators and neutralization antibodies in PLWH. T-cell responses (Th1, Th2, Th17, Treg) and intracellular expression of cytokines (IL-2 and TNF-α) in CD4 or CD8 were induced by spike protein in SARS-CoV-2 (SARS-2-S) and further measured by intracellular staining. ResultsCD4, B cells, CD4+CD45RO+ memory T cells in peripheral blood of PLWH are dramatically decreased in comparison with HD. Importantly, PLWH display comparable neutralizing antibody positive rate to HD after inoculation with inactivated SARS-CoV-2 vaccine. However, PLWH showed weaker responses to vaccines exhibited by lower levels of neutralizing antibodies. Correlation analysis shows that this is possibly caused by low number of CD4 and B cells. Furthermore, SARS-2-S-induced Th2 and Th17 responses are also decreased in PLWH, while no influences on Treg and other cytokines (IL-2, TNF-α and IFN-γ) observed. ConclusionsPLWH and HD have comparable neutralizing antibodies positive rates, but PLWH display weaker responses to inactivated SARS-CoV-2 vaccines in magnitude, which suggests that a booster dose or dose adjustment are required for HIV-infected individuals, especially for those with lower counts of CD4 T and B cells.

Non-infectious outer membrane vesicles derived from Brucella abortus S19Δper as an alternative acellular vaccine protects mice against virulent challenge

The prime human and animal safety issues accentuate the search of promising newer alternative vaccine candidates to resolve complications associated with the live attenuated Brucella abortus strain19 (S19) vaccine. Outer membrane vesicles (OMVs S19 Δper) extracted from Brucella abortus S19Δper (S19Δper) as an alternative subunit vaccine candidate has been explored in the present study as OMVs are endowed with immunogenic molecules, including LPS and outer membrane proteins (OMPs) and do not cause infection by virtue of being an acellular entity. The LPS defective S19Δper released a higher amount of OMVs than its parent strain S19. Under transmission electron microscopy (TEM), OMVs were seen as nano-sized outward bulge from the surface of Brucella. Dynamic light scattering analysis of OMVs revealed that OMVs S19Δper showed the less polydispersity index (PDI) than OMVs S19 pointing towards relatively more homogenous OMVs populations. Both OMVs S19Δper and OMVs S19 with or without booster dose and S19 vaccine were used for immunization of mice and subsequently challenged with 2 × 105 CFU virulent Brucella abortus strain 544 (S544) to assess protective efficacy of vaccines. The less splenic weight index and less S544 count in OMVs immunized mice in comparison to unimmunized mice after S544 challenge clearly indicated good protective efficacy of OMVs. OMVs S19 Δper induced relatively high titer of IgG than OMVs S19 but conferred nearly equal protection against brucellosis. An ELISA based determination of IgG and its isotype response, Cytometric Bead Array (CBA) based quantitation of serum cytokines and FACS based enumeration of CD4+ and CD8+ T cells revealed high titer of IgG, production of both Th1 (IgG2a) and Th2 (IgG1) related antibodies, stimulation of IL-2, TNF (Th1) and IL-4, IL-6, IL-10 (Th2) cytokines, and induced T cell response suggested that OMVs S19Δper elicited Th1 and Th2 type immune response and ensured protection against S544 challenge in murine model.

Performance evaluation of the novel portable PIMA point-of-care CD4 analyzer in comparison with conventional flow cytometer instruments in Cameroon

BackgroundAbsolute CD4+ T-lymphocyte counts are used in the initiation and monitoring of antiretroviral therapy in HIV-infected patients: with the increase number of HIV infected patient and the reduce number of heath care system in rural zones, HIV monitoring in resource-constrained settings demands affordable and reliable CD4+ T lymphocytes enumeration methods. We evaluated a simple PIMA POC which is a dedicated system for enumeration that uses immunomagnetic and immunofluorescent technologies. The instrument was designed to be a low-cost, yet reliable and robust one. In this study, we assessed the correlation between most representative flow cytometry instruments present in Cameroon instead of CyFlow from PARTEC, FACSCount, and FACSCalibur both from Becton Dickinson.MethodsCD4 surface markers on lymphocytes was measured on samples collected in EDTA tubes from 268 patients aged from 1 to 65 years old in three different health care structures. HIV infected patients are coming from CIRCB, Day Hospital of Hopital Central de Yaounde (HCY) and Hopital General de Yaounde (HGY). After inform consent, samples were collected and 101 samples were tested with the FACSCalibur, 60 samples were tested with the CyFlow and 107 samples were tested with the FACSCount flow cytometers. All these samples were tested by different technician with PIMA POC present in all these health care structures and the correlation and agreement were analyzed using linear regression and Bland–Altman analysis.ResultsThe PIMA POC system has excellent precision, accuracy and linearity for CD4+ T lymphocytes enumeration. Good correlations were obtained between the PIMA POC system and other single platform methods. Bland–Altman plots showed interchangeability between the three machines. Absolute CD4+ T-lymphocyte values obtained from the PIMA system correlated well with Cyflow, FACSCount, and FACSCalibur method (r2 varies from 0.88 to 0.968, P < 0.0001). The comparison between values obtained from PIMA with CYFLOW, FACSCount, and FACSCalibur give P = 0.17, P = 0.5 and P = 0.6 respectively meaning that there is not significant differences between values obtained with PIMA and other flow machines.ConclusionThis POC PIMA system is a simple and reliable system for enumeration of absolute CD4+ T-lymphocytes. Having one PIMA system easy to use, should reduce the cost and thus increase access to CD4 testing for HIV infected patients in resource-constrained countries. POC CD4 may also alleviate testing burdens at traditional central CD4 laboratories, hence improving test access in both rural and urban environments. This will reduce also the loss of follow up.

Multi-HLA class II tetramer analyses of citrulline-reactive T cells and early treatment response in rheumatoid arthritis

BackgroundHLA class II tetramers can be used for ex vivo enumeration and phenotypic characterisation of antigen-specific CD4+ T cells. They are increasingly applied in settings like allergy, vaccination and autoimmune diseases. Rheumatoid arthritis (RA) is a chronic autoimmune disorder for which many autoantigens have been described.ResultsUsing multi-parameter flow cytometry, we developed a multi-HLA class II tetramer approach to simultaneously study several antigen specificities in RA patient samples. We focused on previously described citrullinated HLA-DRB1*04:01-restricted T cell epitopes from α-enolase, fibrinogen-β, vimentin as well as cartilage intermediate layer protein (CILP). First, we examined inter-assay variability and the sensitivity of the assay in peripheral blood from healthy donors (n = 7). Next, we confirmed the robustness and sensitivity in a cohort of RA patients with repeat blood draws (n = 14). We then applied our method in two different settings. We assessed lymphoid tissue from seropositive arthralgia (n = 5) and early RA patients (n = 5) and could demonstrate autoreactive T cells in individuals at risk of developing RA. Lastly, we studied peripheral blood from early RA patients (n = 10) and found that the group of patients achieving minimum disease activity (DAS28 < 2.6) at 6 months follow-up displayed a decrease in the frequency of citrulline-specific T cells.ConclusionsOur study demonstrates the development of a sensitive tetramer panel allowing simultaneous characterisation of antigen-specific T cells in ex vivo patient samples including RA ‘at risk’ subjects. This multi-tetramer approach can be useful for longitudinal immune-monitoring in any disease with known HLA-restriction element and several candidate antigens.

Abstract B96: Pharmacodynamic studies of SB 11285, a systemically bioavailable STING agonist in orthotopic tumor models

Abstract Background: The activation of innate and adaptive immunity via Stimulator of Interferon Genes (STING) signaling is a potentially transformative immunotherapeutic strategy in cancer. We have previously reported that SB 11285 is a novel and highly potent STING agonist that can be delivered by intravenous, intraperitoneal and intratumoral routes. We report here the antitumor and pharmacodynamic studies of SB 11285 in multiple orthotopic and subcutaneous syngeneic mouse tumor models. Methods: (1) Orthotopic NBT-II syngeneic rat bladder model: Rat bladder tumor cells were surgically implanted in the bladder wall of rats (n =10). After 6 days, SB 11285 was administered by i.v. @ 0.5, 1.5, and 3mg/kg on days 1, 5, 9,1 3, 17, 21, 25 and 31. Groups of rats treated with docetaxel at 7.5mg/kg i.v. and saline were used as positive and negative controls, respectively. At the end of dosing period, the bladders were harvested and evaluated by measurement of bladder weight and gross pathology. (2) Pharmacodynamic (PD) studies: Syngeneic mouse subcutaneous CT26 colon carcinoma and orthotopic 4T1 breast cancer models were used. In the CT26 model, when mean tumor volume (MTV) reached 125mm3, SB 11285 was administered at 3mg/kg i.v on days 1 and 5. In the orthotopic 4T1 model, SB 11285 was administered i.p. @ 10mg/kg on days 5, 7, 9, 11, 13, 17 and 19. Blood, lymph nodes, spleen and tumor were harvested and evaluated for (i) cytokine analysis by multiplexing assays and (ii) enumeration of CD4+, CD8+, Tregs, macrophages and MDSCs in blood, lymph nodes, spleen and tumor. Results: (1) In the orthotopic NBT-II rat bladder study, animals that received SB 11285 in all doses remained tumor-free based upon gross pathology and bladder weight measurements. SB 11285 was shown to be well tolerated and safe. (2) In both the 4T1 and CT26 PD studies, cytokine analysis of blood and tissues showed strong induction of type I interferon signature without significant systemic inflammatory response. Flow cytometric analysis of blood, lymph nodes, spleen and tumor revealed significant increase in macrophage &amp; CD8+ T-cell infiltration and decrease in Tregs &amp; CD4+ T-cell infiltration. Conclusion: SB 11285 showed potent antitumoral activity in multiple orthotopic and subcutaneous models that correlated with antitumoral immune response mediated by the induction of IFNs, and ISGs and other cytokines along with reduction of Tregs. SB 11285 is being advanced to human clinical trials. Citation Format: Sreerupa Challa, Balaji Ramachandran, Lalitha Vijayakrishnan, Douglas Weitzel, Shenghua Zhou, Kris Iyer. Pharmacodynamic studies of SB 11285, a systemically bioavailable STING agonist in orthotopic tumor models [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr B96.

Issue Highlights - May 2019.

Issue Highlights - May 2019.

CD4+ CD25+ FOXP3+ T cell frequency in the peripheral blood is a biomarker that distinguishes intestinal tuberculosis from Crohn's disease.

BackgroundDistinguishing between Crohn’s Disease (CD) and Intestinal Tuberculosis (ITB) has been a challenging task for clinicians due to their similar presentation. CD4+FOXP3+ T regulatory cells (Tregs) have been reported to be increased in patients with pulmonary tuberculosis. However, there is no such data available in ITB. The aim of this study was to investigate the differential expression of FOXP3+ T cells in patients with ITB and CD and its utility as a biomarker.MethodsThe study prospectively recruited 124 patients with CD, ITB and controls: ulcerative colitis (UC) and patients with only haemorrhoidal bleed. Frequency of CD4+CD25+FOXP3+ Tregs in peripheral blood (flow cytometry), FOXP3 mRNA expression in blood and colonic mucosa (qPCR) and FOXP3+ T cells in colonic mucosa (immunohistochemistry) were compared between controls, CD and ITB patients.ResultsFrequency of CD4+CD25+FOXP3+ Treg cells in peripheral blood was significantly increased in ITB as compared to CD. Similarly, significant increase in FOXP3+ T cells and FOXP3 mRNA expression was observed in colonic mucosa of ITB as compared to CD. ROC curve showed that a value of >32.5% for FOXP3+ cells in peripheral blood could differentiate between CD and ITB with a sensitivity of 75% and a specificity of 90.6%.ConclusionPhenotypic enumeration of peripheral CD4+CD25+FOXP3+ Treg cells can be used as a non-invasive biomarker in clinics with a high diagnostic accuracy to differentiate between ITB and CD in regions where TB is endemic.

Rapid enumeration of CD4 + T lymphocytes using an integrated microfluidic system based on Chemiluminescence image detection at point-of-care testing.

An integrated microfluidic system has been developed for rapid enumeration of CD4 + T lymphocytes at point-of-care (POC) settings. A concise microfluidic chip, which consists of three separate chambers, respectively, for reaction, detection and waste storage, is developed to automate CD4 detection. To simplify CD4 + T lymphocyte enumeration, a single polycarbonate bead immobilized with CD4 antibody is adopted by the microfluidic chip to capture the CD4 antigen in the lysed testing sample. Desired performance is achieved by actuating the single bead for efficient mixing, as well as transferring it between different reaction chambers to reduce non-specific reaction. A controllable external magnetic field is applied to drive the single bead with a built-in ferrous core for different purposes. Chemiluminescence reaction is implemented in an independent chamber to reduce non-specific binding of enzyme. A simple flow control strategy is adopted to conveniently release the waste reagent into the waste storage chamber by just opening the vent hole without actively pumping. A sensitive CCD camera is used to collect the reaction signal by taking picture from the single bead, and then the signal intensity is further analyzed for CD4 + T lymphocyte enumeration. Experimental results show that rapid, convenient, accurate and low-cost CD4 + T lymphocyte enumeration can be obtained with the developed microfluidic system at POC test.

Human Immuno-Deficiency Virus Co-Infection with Hepatitis B Virus and Baseline CD4+ T Cell Count among Patients Attending a Tertiary Care Hospital, Nepal

Background: Since 1981, when the first AIDS case was reported, worldwide, more than 34 million people have been infected with HIV. Almost 95 percent of the people infected with HIV live in developing countries. As HBV &amp; HIV share similar routes of transmission by sexual intercourse or drug use by parenteral injection, co-infection is common. Because of the limited access to healthcare &amp; HIV treatment in developing countries, HIV-infected individuals are present late for care. Enumeration of CD4+ T cell count at the time of diagnosis has been useful to initiate the therapy in HIV infected individuals. The baseline CD4+ T cell count shows high immunological variability among patients. Methods: This prospective study was done in the serology section of the Department of Microbiology over a period of one year from august 2012 to July 2013. A total of 13037 individuals subjected for HIV test were included in the study comprising of 4982 males &amp; 8055 females. Blood sample was collected by vein puncture aseptically with standard operational procedure in clean &amp; dry test-tube. All blood samples were screened for HIV as described by WHO algorithm by Immuno-chromatography rapid kits. Further confirmation was done by biokit ELISA method as per the manufacturer’s guidelines. After informed consent, HIV positive individuals were screened for HBsAg by Immuno-chromatography rapid kits (Hepacard). Further confirmation was done by biokit ELISA method as per the manufacturer’s guidelines. EDTA blood samples were collected from the HIV sero-positive individuals for baseline CD4+ T count. Then, CD4+ T cells count was determined by using FACS Calibur Flow Cytometer (BD). Results: Among 13037 individuals screened for HIV, 104 (0.8%) were found to be infected comprising of 69(66.34%) males &amp; 35 (33.65%) females. The study showed that the high infection was noted in active age group (30.76%), housewives (28.7%) &amp; in heterosexual route (80.9%) of transmission. Out of total HIV infected individuals, distribution of HBV co-infection was found to be 6 (5.7%). Baseline CD4+ T cell count of HIV infected patient was found higher (mean CD4+ T cell count; 283cells/cu.mm) than HBV co infected patients (mean CD4+ T cell count; 91 cells/cu.mm). Majority (77.2%) of HIV infected &amp; all co-infected individuals were presented in our center late (CD4+ T cell count;&lt; 350/cu. mm) for diagnosis and care. Majority of co- infected 4 (80%) were late presented with advanced AIDS stage (CD4+ count; &lt;200/cu.mm). Conclusions: The study showed a high percentage of HIV sero-positive &amp; co- infected individuals. Baseline CD4+ T cell count of majority of HIV infected individuals was found to be low. Hence, more sustained and vigorous awareness campaigns &amp; counseling still need to be done in order to promote early diagnosis and management.

CORRELATION OF HIV-1 P24 ASSAY WITH CD4 T-CELL COUNT, HIV, HBV AND HCV CO-INFECTIONS AND ITS IMPLICATION FOR ART MONITORING IN VASTLY HIV-INFECTED POPULATION OF NIGERIA

BackgroundReports indicate that extensive genetic diversity of HIV-1 impacts almost every aspect of HIV-1 epidemiology, including laboratory detection, ART/resistance, monitoring of ART and vaccine development. Therefore, in order to support...

Enumeration of CD4 + T lymphocytes Count among Healthy HIV Seronegative Individuals & People Living with HIV in Manipur

Introduction: The CD4+ T lymphocytes are the primary target cells for Human Immunodeficiency Virus (HIV). The enumeration of CD4+ T cells is the most important marker for HIV disease progression. The estimation of CD4+ T cell counts is used to decide the initiation of anti retroviral therapy (ART), to monitor the efficacy and to start treatment for opportunistic infections (OIs). Hence it is important to establish the reference ranges for the CD4+ T cell counts in the target population to understand the immune dysfunction.Objectives: To determine the reference range of CD4 + T cell counts in healthy HIV seronegative individuals and its comparison with those people living with HIV in Manipur.Materials and Methods: The study was carried out between August 2013 to June 2015 in the Dept. Of Microbiology, RIMS, Imphal, Manipur. Blood samples from 334 HIV seronegative volunteers comprising group I (188 males and 146 females) and 1058 HIV-positive patients comprising group II (525 males and 533 females) in the age group of 17-60 years, were analyzed for enumeration of CD4+ T cell counts by fluorescent activated cell sorter (FACS) counter (Becton Dickinson).Results: In group I mean CD4 levels were 741 180.349 cells/l. In normal healthy males CD4 levels were 656.99 122.73 cells/l and 849.36 185.190 cells/l among healthy females. In group II mean CD4 levels were 341.43 301.885 cells/l. Among the HIV positive males CD4 levels were 300.35 200.634 cells/l while among the females CD4 levels were 381.89 371.638 cells/l.Conclusion: As there is heterogeneity in baseline CD4 + T cell counts, our findings on T-cell subset reference ranges of normal healthy seronegative people in Manipur validate the utility of determination of CD4 cell count.

Evaluation of the FACSPresto, a New Point of Care Device for the Enumeration of CD4% and Absolute CD4+ T Cell Counts in HIV Infection.

IntroductionEnumeration of CD4+ T lymphocytes is important for pre-ART disease staging and screening for opportunistic infections, however access to CD4 testing in resource limited settings is poor. Point of care (POC) technologies can facilitate improved access to CD4 testing. We evaluated the analytical performance of a novel POC device the FACSPresto compared to the FACSCalibur as a reference standard and to the PIMA, a POC device in widespread use in sub-Saharan Africa.MethodSpecimens were obtained from 253 HIV infected adults. Venous blood samples were analyzed on the FACSPresto and the FACSCalibur, in a subset of 41 samples additional analysis was done on the PIMA.ResultsThe absolute CD4 count results obtained on the FACSPresto were comparable to those on the FACSCalibur with low absolute (9.5cells/μl) and relative bias (3.2%). Bias in CD4% values was also low (1.06%) with a relative bias of 4.9%. The sensitivity was lower at a CD4 count threshold of ≤350cells/μl compared with ≤500cells/μl (84.9% vs. 92.8%) resulting in a high upward misclassification rate at low CD4 counts. Specificity at thresholds of ≤350cells/μl and ≤500cells/μl were 96.6% and 96.8% respectively. The PIMA had a high absolute (-68.6cells/μl) and relative bias (-10.5%) when compared with the FACSCalibur. At thresholds of ≤350cells/μl and ≤500cells/μl the sensitivity was 100% and 95.5% respectively; specificity was 85.7% and 84.2% respectively. The coefficients of repeatability were 4.13%, 5.29% and 9.8% respectively.DiscussionThe analytic performance of the FACSPresto against the reference standard was very good with better agreement and precision than the PIMA. The FACSPresto had comparable sensitivity at a threshold of 500 cells/μl and better specificity than the PIMA. However the FACSPresto showed reduced sensitivity at low CD4 count thresholds.ConclusionThe FACSPresto can be reliably used as a POC device for enumerating absolute CD4 count and CD4% values.

PIMA™ point-of-care testing for CD4 counts in predicting antiretroviral initiation in HIV-infected individuals in KwaZulu-Natal, Durban, South Africa.

IntroductionLimited information is available on the usefulness of the PIMA™ analyser in predicting antiretroviral treatment eligibility and outcome in a primary healthcare clinic setting in disadvantaged communities in KwaZulu-Natal, South Africa.Materials and methodsThe study was conducted under the eThekwini Health Unit, Durban, KwaZulu-Natal. Comparison of the enumeration of CD4+ T-cells in 268 patients using the PIMA™ analyser and the predicate National Health Laboratory Services (NHLS) was undertaken during January to July 2013. Bland-Altman analysis to calculate bias and limits of agreement, precision and levels of clinical misclassification at various CD4+ T-cell count thresholds was performed.ResultsThere was high precision of the PIMA™ control bead cartridges with low and normal CD4+ T-cell counts using three different PIMA™ analysers (%CV < 5). Under World Health Organization (WHO) guidelines (≤ 500 cells/mm3), the sensitivity of the PIMA™ analyser was 94%, specificity 78% and positive predictive value (PPV) 95%. There were 24 (9%) misclassifications, of which 13 were false-negative in whom the mean bias was 149 CD4+ T-cells/mm3. Most (87%) patients returned for their CD4 test result but only 67% (110/164) of those eligible (≤ 350 cells/mm3) were initiated on antiretroviral therapy (ART) with a time to treatment of 49 days (interquartile range [IQR], 42–64 days).ConclusionThere was adequate agreement between PIMA™ analyser and predicate NHLS CD4+ T-cell count enumeration (≤ 500 cells/mm3) in adult HIV-positive individuals. The high PPV, sensitivity and acceptable specificity of the PIMA™ analyser technology lend it as a reliable tool in predicting eligibility and rapid linkage to care in ART programmes.

CD4 cells profile of HAART naive HIV seropositive clients in Kogi State University Teaching Hospital, Anyigba, Kogi State. Nigeria

CD4 lymphocyte cells are the primary targets of Human immune-deficiency virus (HIV). Enumeration of CD4 T lymphocytes in the peripheral blood is used in the assessment of disease clinical stage, risk of opportunistic infections, evaluation of prognosis and guide decision on the commencement of antiretroviral therapy. The objective was to determine CD4 cells profile of HIV sero-positive naive patients in Kogi State University Teaching Hospital( KSUTH) Anyigba. A total of 404 HIV sero-positive Highly Active Anti Retro Viral Treatment ( HAART) naive patients comprising 147(36.4%) males and 257 (63.6%) females were examined. Approval was obtained from ethical committee of Kogi State University Teaching Hospital (KSUTH), Anyigba. Written and verbal informed consent was taken from all patients. The overall mean age of patients was 33.0 ± 12.7 years and female-male ratio was 1.7:1. Majority of patients were in the clinical stage two 121(30.5%) and three 200(50.4%). Patients had overall mean CD4 cells count of 381.8 ± 240.8 cells /mm3. Patients CD4 cells count varied statistically with the HIV clinical staging (F =4.512 & P value=0.004) and statistically insignificant with gender (P value = 0.7562 & t Test= 0.3106) and tuberculosis status (P value=0.223 & F= 1.505).Conclusion: This study showed HIV sero-positive HAART naive patients presented in KSUTH with mean age of 33 years. Majority of patients presented in disease clinical stage two and three with a mean CD4 cells counts of 381.8 cells/mm3 . This study recommend the need to reduce stigmatization, discrimination and promote early access to treatment , care and support services. Keywords: HIV/AIDS, CD4, Patient/clients, KSUTH, Nigeria.

Triple positivity of HBsAg, anti-HCV antibody, and HIV and their influence on CD4+ lymphocyte levels in the highly HIV infected population of Abeokuta, Nigeria.

Few studies exist on hospital-based seroprevalence of triple positivity of HIV/HBV/HCV in Nigeria. The study aimed at determining the triple positivity of HIV, HBsAg and HCV among HIV-infected individuals in Abeokuta, Nigeria and defining the influence of these triple infections on CD4+ counts of HIV-infected individuals as antiretroviral therapy improves in Nigeria. Enumeration of CD4+ levels in 183 HIV-infected persons was done with Partec Flow Cytometer. Seropositivity of HBsAg and anti-HCV antibody was detected with rapid kits. From the result obtained, significance variance (p<0.05) existed between HIV positive persons and persons who tested positive to HIV/HBV/HCV triple infection before and after the commencement of HAART. Of these infections, 31(16.9%) had HBV/HCV/HIV triple infection, while 152(83.1%) had HIV mono infection only, 56(30.6%) had HBV/HIV dual infection only and 43(23.5%) had HCV/HIV dual infection only. Significant variance (p<0.05) also existed between subjects with CD4 counts of <200 cells/µl, 200-499 cells/µl and >500 cells/µl. Highest seroprevalence of HIV (35.0%) was found in age groups 35-44 years and >65 years had the least (2.7%). Significant variance (p<0.05) also existed in the progression of CD4+ lymphocytes cells between subjects with persistent decrease (32.3%) in CD4+ lymphocytes cells and those with fluctuation in their CD4+ lymphocytes cells (12.9%) after the commencement of ART. The study further confirms that triple positivity of HIV/HBV/HCV infection is common in Abeokuta, Nigeria. Testing of these triple infections should be a big concern in the best choice and commencement of ART. Also, the study showed that consistent and prolonged use of HAART had a positive impact on the CD4 count of HIV-infected individuals.

Issue Highlight 2015.

Issue Highlight 2015.