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Indian Journal of Medical Microbiology, Vol. 28, No. 2, April-June, 2010, pp. 183-184 Correspondence Normal CD4 and CD3 lymphocyte counts in healthy south Indian adults *S Srirangaraj, D Venkatesha Department of Microbiology (SS), Mahatma Gandhi Medical College and Research Institute, Pondicherry - 605 012, Department of Microbiology (DV), Mysore Medical College and Research Institute, Mysore - 570 001, Karnataka, India Correspondence Address: Department of Microbiology, Mahatma Gandhi Medical College and Research Institute, Pondicherry - 605 012, India, rangaraj1980@indiatimes.com Date of Submission: 22-Sep-2009 Code Number: mb10056 PMID: 20404475 DOI: 10.4103/0255-0857.62506 Dear Editor, CD4 + T-cell enumeration is an important surrogate marker for HIV disease progression. Flow cytometry is the gold standard technique for CD4 enumeration. The information on normal CD4 counts from different regions of the country will help establish the normal ranges of the CD4 counts in our population. [1] This in turn may be of help in deciding whether the western standards of anti-retroviral therapy initiation and management can be used without any changes in the Indian context. This study was undertaken to determine the reference ranges of CD4 and CD3 counts in normal healthy adults in Mysore, India. It was carried out from August 20, 2006 to August 19 2007. We have an integrated counselling and testing centre (ICTC) for HIV and a centre to estimate CD4 /CD3 counts under NACO (National AIDS Control Organisation). The study was conducted after obtaining ethical clearance from the ethics committee of our institution. Informed consent was obtained from each study subject. Fifty healthy individuals above 18 years of age, from both sexes, were included in the study. The selected individuals included resident doctors, medical students, laboratory staff, nurses, blood donors and healthy volunteers. Pre-test and post-test counselling was given to them after obtaining their informed consent for HIV testing. A pre-designed proforma was used to collect socio-demographic and clinical data. Strict confidentiality was maintained. About 5ml venous blood was collected between 9 am to 12 noon following universal precautions, using a 2ml K3-EDTA Vacutainer (BD) for CD4 testing and another 3ml for HIV testing using a plastic vial. The testing for HIV antibodies was done at our ICTC centre using ENZAIDS ELISA kit (Span Diagnostics Ltd., Surat, India), strictly following the manufacturer's instructions. The CD4/CD3 enumeration was done using the single -platform BD FACS Calibur TM machine (Becton, Dickinson and Company, San Jose, United States of America), using the fluorochrome-labelled monoclonal antibodies to CD4 + T-cells and CD3 + T-cells. [2] A TruCount tube (Becton, Dickinson and Company, San Jose, United States of America) was labelled with the sample identification number; 20ìL of TriTEST CD3/CD4/CD45 reagent (Becton, Dickinson and Company, San Jose, United States of America) was pipetted into the bottom of the tube and 50 µL of well- mixed anticoagulated blood was pipetted into the bottom of the tube following the reverse pipetting technique onto the side of the tube just above the retainer, using the micropipettor (Pipetman® , Rainin Instrument Co Inc, USA). The pre-acquisition software set-up was done by launching BDMultiSET software (Becton, Dickinson and Company, San Jose, United States of America). The operator's name was entered in the operator field and 'ACCEPT' was clicked in the set-up view. Entry of sample and panel information was done. The rack was loaded with sample tubes and lymphocyte data acquisition and analysis was done using BDMultiSET software package. Internal quality control was performed with process controls using the manufacturer's recommendations. External quality control was performed through an external quality assurance programme with NARI (National AIDS Research Institute), Pune. SPSS software evaluation version 14 (SPSS Inc. New York, USA) was used for data compilation and analysis. Statistical analysis was done using Chi-square test, 't'test for equality of means, ANOVA and by calculation of mean and standard deviation. The age wise distribution of CD4 counts is given in [Table - 1]. The mean CD4 counts in the present study were 992.52 ± 323 cells/mL. This is comparable with the study from Vellore, where 1048 ± 210 cells/ mL was reported. [3] In contrast, the CD4 counts reported from New Delhi (703 ± 225 cells/mL), Manipal (835±222 cells/mL) and Pune (865 cells/mL, SD not reported) were lower when compared with our study. [1],[4],[5] This shows the regional diversity in the CD4 counts. In our study, the CD4 % was 37.48 ± 0.8 The mean CD3 counts were 1846.76 ± 465.09 cells and the mean CD3 % was 69.19 ± 1.02. The normal CD4 range was 482 cells/µL to 1798 cells/mL. The mean CD4 counts were higher in females than in males. However, this was statistically insignificant. Similar findings were reported by Ray et al. [1] No statistical difference was observed in the CD3 and CD4 counts of smokers and alcoholics when compared with those of teetotallers, which compares with that of Uppal et al. [5] However, the CD45 counts in the present study were higher in alcoholics compared with non-alcoholics (P < 0.05). The significance of this finding is unclear. Acknowledgment The authors are grateful to Mr. Khan Ghori for his technical support. References
Copyright 2010 - Indian Journal of Medical Microbiology The following images related to this document are available:Photo images[mb10056t1.jpg] |
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