Establishment of CD4 and CD8 Lymphocyte subsets in a healthy HIV and Toxoplasma seronegative pregnant women in Libya
Volume 1 Issue: 1 February 2021

Supplementary Files

Download the PDF


CD4 -
CD8 T-lymphocyte counts
Flow Cytometry

How to Cite

Amro, A., Gashout, A., Erhuma, M., Al-Dwibe, H., Almgrhe, A., & Abudaher, A. (2021). Establishment of CD4 and CD8 Lymphocyte subsets in a healthy HIV and Toxoplasma seronegative pregnant women in Libya . Al-Quds University Journal for Academic Research, 1(1). Retrieved from


Most of the diagnostic laboratories in Libya often depend on western textbooks for CD4+- and CD8+ T-lymphocyte reference values. In this paper, we established reference ranges for the Libyan Toxoplasma, HIV, HBV, and HCV seronegative healthy pregnant women in all trimesters of pregnancy, and compared them with a control group of non-pregnant women. Wholeblood samples were collected to provide normal ranges for CD4+ and CD8+ Lymphocyte subsets expressed as mean ± standard division. A total of 110 Libyan women who came from Tripoli and Zwara districts were investigated; 70 pregnant women

(aged 27.8 ± 2.99, range 18-40 years old) and 40 non-pregnant women (aged 22.7±3.01, range 18-40 years old) were included as controls. All cases/controls were seronegative for toxoplasmosis, HIV, HBV and HCV. The CD4+ cell counts were 685±256 cell/#l at the first trimester (T1), 740±202 at T2, and 923±203 cell/#l at T3. While the CD8+ cell counts were 451±171 cell/#l at T1, 541±168 at T2, and 753±190 cell/#l at T3. The CD4:CD8 ratios were 1.5±0.64 at T1, 1.4±0.51 at T2, and 1.2±0.36 at T3. Moreover, the mean absolute CD4+ and CD8+ counts for the control group were 1001±232 cell/#l and 717±159 cell/#l respectively. Absolute counts of CD4+ and CD8+ cells in pregnant women were significantly lower as compared to controls (P<0.05). Statistically significant decrease in the CD4+ and CD8+ cell counts was reported during T1 (P<0.05). These values increased significantly during the T2, and was comparable to the controls during T3 (P>0.05). The absolute CD4+ and CD8+ cell counts

decreased with age for both groups. Geographical variation was reported for the cell counts between Tripoli and Zwara district at T3. We established reference ranges of CD4+ and CD8+ T-lymphocytes for the Libyan healthy pregnant women and discussed their use as prognostic markers. Further cohorts with greater sample size may be required to define the stage of the disease in relation to the normal CD4+ and CD8+ T lymphocyte count subsets in the Libyan population.

Keywords: CD4+-, CD8+ T-lymphocyte counts, Flow Cytometry, Libya, pregnancy, HIV, toxoplasmosis.


Aagaard-Tillery, K. M., R. Silver and J. Dalton (2006). "Immunology of normal pregnancy." Semin Fetal Neonatal Med 11(5): 279-295.

Abimiku, A., P. Villalba-Diebold, J. Dadik, F. Okolo, E. Mang and M. Charurat (2009). "Risk factors associated with low CD4+ lymphocyte count among HIV-positive pregnant women in Nigeria." Int J Gynaecol Obstet 106(3): 227-231.

Aina, O., J. Dadik, M. Charurat, P. Amangaman, S. Gurumdi, E. Mang, R. Guyit, N. Lar, P. Datong, C. Daniyam, P. Kanki and A. Abimiku (2005). "Reference values of CD4 T lymphocytes in human immunodeficiency virusnegative adult Nigerians." Clin Diagn Lab Immunol 12(4): 525-530.

Akinbami, A. A., A. Gbadegesin, S. O. Ajibola, E. I. Uche, A. O. Dosunmu, A. Adediran and A. Sobande (2015). "Factors influencing CD4 cell count in HIV-positive pregnant women in a secondary health center in Lagos, Nigeria." HIV AIDS (Auckl) 7: 115-118.

Alkhunfas, S. R. (2008). "Toxoplasmosis in Newborn Babies in AL-Jala Maternity Hospital, Tripoli – Libya." Master thesis, Biology department, Academy of graduate study –Libya: 45-46.

Bosire, E. M., A. K. Nyamache, M. M. Gicheru, S. A. Khamadi, R. W. Lihana and V. Okoth (2013). "Population specific reference ranges of CD3, CD4 and CD8 lymphocyte subsets among healthy Kenyans." AIDS Res Ther 10(1): 24.

Carmichael, K. F. and A. Abayomi (2006). "Analysis of diurnal variation of lymphocyte subsets in healthy subjects in the Caribbean, and its implication in HIV monitoring and treatment." Afr J Med Med Sci 35(1): 53-57.

Chng, W. J., G. B. Tan and P. Kuperan (2004). "Establishment of adult peripheral blood lymphocyte subset reference range for an Asian population by singleplatform flow cytometry: influence of age, sex, and race and comparison with other published studies." Clin Diagn Lab Immunol 11(1): 168-173.

Dayama, A., D. Pandit, S. Mudaliar, R. Bharadwaj, K. E. Bharucha, A. N. Shrotri and S. Joshi (2003). "A pilot study on CD4 & CD8 cell counts in healthy HIV seronegative pregnant women." Indian J Med Res 117: 198-200.

Dubey, J. P. and J. L. Jones (2008). "Toxoplasma gondii infection in humans and animals in the United States." Int J Parasitol 38(11): 1257-1278.

Fahey, J. L., J. F. Schnelle, J. Boscardin, J. K. Thomas, M. E. Gorre, N. Aziz, H. Sadeghi and P. Nishanian (2000). "Distinct categories of immunologic changes in frail elderly." Mech Ageing Dev 115(12): 1-20.

Feldman, J. G., H. Minkoff, M. F. Schneider, S. J. Gange, M. Cohen, D. H. Watts, M. Gandhi, R. S. Mocharnuk and K. Anastos (2006). "Association of cigarette smoking with HIV prognosis among women in the HAART era: a report from the women's interagency HIV study." Am J Public Health 96(6): 1060-1065.

Force, E. C. a. U. S. P. H. S. T. (1994). "Recommendations of the Us-Public-Health-Service Task-Force on the Use of Zidovudine to Reduce Perinatal Transmission of Human-ImmunodeficiencyVirus (Reprinted from Mmwr, Vol 43, Pg 1-20, 1994)." Pediatric Aids and Hiv Infection-Fetus to Adolescent 5(6): 387-399.

Gashout, A., A. Amro, M. Erhuma, H. Al-Dwibe, E. Elmaihub, H. Babba, N. Nattah and A. Abudher (2016). "Molecular diagnosis of Toxoplasma gondii infection in Libya." BMC Infect Dis 16: 157.

Gashout, A., T. Lazrag, H. Gashout and T. Swedan (2008). "Qualtitive assessement of risk for spotaneous abortion associated with toxoplasma and rubella: immunity appraisal." The Libyan Journal of Infectious Disease 2(1): 52-56.

Greve, B., U. Cassens, C. Westerberg, W. G. Jun, W. Sibrowski, D. Reichelt and W. Gohde (2003). "A new no-lyse, no-wash flow-cytometric method for the determination of CD4 T cells in blood samples." Transfusion Medicine and Hemotherapy 30(1): 8-13.

Halonen, S. K. and L. M. Weiss (2013). "Toxoplasmosis." Handb Clin Neurol 114: 125-145.

Hide, G., O. Gerwash, E. K. Morley, R. H. Williams, J. M. Hughes, D. Thomasson, M. S. Elmahaishi, K. H. Elmahaishi, R. S. Terry and J. E. Smith (2007). "Does vertical transmission contribute to the prevalence of toxoplasmosis?" Parassitologia 49(4): 223-226.

Hide, G., E. K. Morley, J. M. Hughes, O. Gerwash, M. S. Elmahaishi, K. H. Elmahaishi, D. Thomasson, E. A. Wright, R. H. Williams, R. G. Murphy and J. E. Smith (2009). "Evidence for high levels of vertical transmission in Toxoplasma gondii." Parasitology 136(14): 1877-1885.

Hunt, J. S. (2006). "Stranger in a strange land." Immunol Rev 213: 36-47.

Ibitokou, S., L. Brutus, B. Vianou, M. Oesterholt, A.

Massougbodji, P. Deloron, M. Troye-Blomberg, N. Fievet and A. J. Luty (2013). "Gestational agerelated changes in the peripheral blood cell composition of sub-Saharan African women." J Reprod Immunol 98(1-2): 21-28.

Janeway Jr, C. A., P. Travers, M. Walport and M. J. Shlomchik (2001). The components of the immune system. Immunobiology: The Immune System in Health and Disease. 5th edition, Garland Science.

Jenum, P. A. and B. Stray-Pedersen (1998). "Development of specific immunoglobulins G, M, and A following primary Toxoplasma gondii infection in pregnant women." J Clin Microbiol 36(10): 2907-2913.

Jiang, W., L. Kang, H. Z. Lu, X. Pan, Q. Lin, Q. Pan, Y. Xue, X. Weng and Y. W. Tang (2004). "Normal values for CD4 and CD8 lymphocyte subsets in healthy Chinese adults from Shanghai." Clin Diagn Lab Immunol 11(4): 811-813.

Jones, J., A. Lopez and M. Wilson (2003). "Congenital toxoplasmosis." Am Fam Physician 67(10): 21312138.

Kassem, H. H. and T. A. Morsy (1991). "The prevalence of antiToxoplasma antibodies among pregnant women in Benghazi, (S.P.L.A.J.) Libya." J Egypt Soc Parasitol 21(1): 69-74.

Landay, A. L. and K. A. Muirhead (1989). "Procedural guidelines for performing immunophenotyping by flow cytometry." Clin Immunol Immunopathol 52(1): 48-60.

Lebranchu, Y., G. Thibault, D. Degenne and P. Bardos (1991). "Abnormalities in CD4+ T lymphocyte subsets in patients with common variable immunodeficiency." Clin Immunol Immunopathol

(1): 83-92.

Lee, B. W., H. K. Yap, F. T. Chew, T. C. Quah, K. Prabhakaran, G. S. Chan, S. C. Wong and C. C. Seah (1996). "Age- and sex-related changes in lymphocyte subpopulations of healthy Asian subjects: from birth to adulthood." Cytometry 26(1): 8-15.

Levin, A., G. Brubaker, J. S. Shao, D. Kumby, T. R. O'Brien, J. J. Goedert, K. W. Strauss, W. A. Blattner and I. Hannet (1996). "Determination of T-lymphocyte subsets on site in rural Tanzania: results in HIV-1 infected and non-infected individuals." Int J STD AIDS 7(4): 288291.

Liu, Q., Z. D. Wang, S. Y. Huang and X. Q. Zhu (2015). "Diagnosis of toxoplasmosis and typing of Toxoplasma gondii." Parasit Vectors 8: 292.

Lugada, E. S., J. Mermin, F. Kaharuza, E. Ulvestad, W. Were, N. Langeland, B. Asjo, S. Malamba and R.

Downing (2004). "Population-based hematologic and immunologic reference values for a healthy Ugandan population." Clin Diagn Lab Immunol

(1): 29-34.

Magrhi, S., A. Abudher, N. Guma, H. Hagrasi, S. Mohammed, M. Musbah, M. Ali and S. Abeed (2003). "Toxoplasmosis and Pregnancy outcome." Second national Biotechnology, Conference proceeding Book: 435-442.

Mair, C., S. E. Hawes, H. D. Agne, P. S. Sow, I. N'Doye, L. E. Manhart, P. L. Fu, G. S. Gottlieb and N. B. Kiviat (2008). "Factors associated with CD4 lymphocyte counts in HIV-negative Senegalese individuals." Clin Exp Immunol 151(3): 432-440.

McIntire, R. H. and J. S. Hunt (2005). "Antigen presenting cells and HLA-G--a review." Placenta 26 Suppl A: S104-109.

Montoya, J. G. and O. Liesenfeld (2004). "Toxoplasmosis." Lancet 363(9425): 1965-1976.

Montoya, J. G. and J. S. Remington (2008). "Management of Toxoplasma gondii infection during pregnancy." Clin Infect Dis 47(4): 554-566.

Mousa, D. A., M. A. Mohammad and A. B. Toboli (2011). "Toxoplasma gondii infection in pregnant women with previous adverse pregnancy outcom." Medical Journal of Islamic World Academy of Sciences 19(2): 95-102.

Munoz-Suano, A., A. B. Hamilton and A. G. Betz (2011). "Gimme shelter: the immune system during pregnancy." Immunol Rev 241(1): 20-38.

Murugavel, K. G., P. Balakrishnan, J. Mohanakrishnan, S.

S. Solomon, E. M. Shankar, S. P. Muthu Sundaram,

N. Kumarasamy, E. Piwowar-Manning, E. Livant, K. H. Mayer, S. P. Thyagarajan and S. Solomon (2009). "Establishment of T-lymphocyte subset reference intervals in a healthy adult population in Chennai, India." Indian J Med Res 129(1): 59-63.

Ortiz-Alegria, L. B., H. Caballero-Ortega, I. CanedoSolares, C. P. Rico-Torres, A. Sahagun-Ruiz, M. E. Medina-Escutia and D. Correa (2010). "Congenital toxoplasmosis: candidate host immune genes relevant for vertical transmission and pathogenesis." Genes Immun 11(5): 363-373.

Roberts, A., K. Hedman, V. Luyasu, J. Zufferey, M. H. Bessieres, R. M. Blatz, E. Candolfi, A. Decoster,

G. Enders, U. Gross, E. Guy, M. Hayde, D. Ho-Yen,

J. Johnson, B. Lecolier, A. Naessens, H. Pelloux, P. Thulliez and E. Petersen (2001). "Multicenter evaluation of strategies for serodiagnosis of primary infection with Toxoplasma gondii." Eur J Clin Microbiol Infect Dis 20(7): 467-474.

Schaberg, T., C. Theilacker, O. T. Nitschke and H. Lode (1997). "Lymphocyte subsets in peripheral blood

and smoking habits." Lung 175(6): 387-394.

Sun, X., H. Lu, B. Jia, Z. Chang, S. Peng, J. Yin, Q. Chen and N. Jiang (2013). "A comparative study of Toxoplasma gondii seroprevalence in three healthy Chinese populations detected using native and recombinant antigens." Parasit

Vectors 6(1): 241.

Temmerman, M., N. Nagelkerke, J. Bwayo, E. N. Chomba, J. Ndinya-Achola and P. Piot (1995). "HIV-1 and immunological changes during pregnancy: a comparison between HIV-1-seropositive and HIV-1seronegative women in Nairobi, Kenya." AIDS 9(9): 1057-1060.

Tenter, A. M., A. R. Heckeroth and L. M. Weiss (2000). "Toxoplasma gondii: from animals to humans." Int J Parasitol 30(12-13): 1217-1258.

Tollerud, D. J., J. W. Clark, L. M. Brown, C. Y. Neuland, L. K. Pankiw-Trost, W. A. Blattner and R. N. Hoover (1989). "The influence of age, race, and gender on peripheral blood mononuclear-cell subsets in healthy nonsmokers." J Clin Immunol 9(3): 214222.

Tsegaye, A., T. Messele, T. Tilahun, E. Hailu, T. Sahlu, R. Doorly, A. L. Fontanet and T. F. Rinke de Wit (1999). "Immunohematological reference ranges for adult Ethiopians." Clin Diagn Lab Immunol 6(3): 410-414.

Tugume, S. B., E. M. Piwowar, T. Lutalo, P. N. Mugyenyi,

R. M. Grant, F. W. Mangeni, K. Pattishall and E. Katongole-Mbidde (1995). "Hematological reference ranges among healthy Ugandans." Clin Diagn Lab Immunol 2(2): 233-235.

Tuomala, R. E., L. A. Kalish, C. Zorilla, H. Fox, W. Shearer,

A. Landay, S. H. Vermund, S. Landesman and D. Burns (1997). "Changes in total, CD4+, and CD8+ lymphocytes during pregnancy and 1 year postpartum in human immunodeficiency virus-infected women. The Women and Infants Transmission Study." Obstet Gynecol 89(6): 967974.

Uppal, S. S., S. Verma and P. S. Dhot (2003). "Normal values of CD4 and CD8 lymphocyte subsets in healthy indian adults and the effects of sex, age, ethnicity, and smoking." Cytometry B Clin Cytom 52(1): 32-36.

Vassiliadou, N. and J. N. Bulmer (1998). "Characterization of endometrial T lymphocyte subpopulations in spontaneous early pregnancy loss." Hum Reprod

(1): 44-47.

Wegmann, T. G., H. Lin, L. Guilbert and T. R. Mosmann (1993). "Bidirectional cytokine interactions in the maternal-fetal relationship: is successful pregnancy a TH2 phenomenon?" Immunol Today

(7): 353-356.

Xiao, Y., J. Yin, N. Jiang, M. Xiang, L. Hao, H. Lu, H. Sang, X. Liu, H. Xu, J. Ankarklev, J. Lindh and Q. Chen (2010). "Seroepidemiology of human Toxoplasma gondii infection in China." BMC Infect Dis 10: 4.

Yang, H., Y. Wu, H. Li, Y. Gao, L. Qu, J. Yang and B. Tao (2015). "Reference values of T lymphocyte subsets among health adults in Inner Mongolia Region." Int J Clin Exp Med 8(3): 4250-4256.

Zhou, Y., Y. Zhang, J. P. Moorman, Z. Q. Yao and Z. S. Jia (2014). "Viral (hepatitis C virus, hepatitis B virus, HIV) persistence and immune homeostasis." Immunology 143(3): 319-330.