Haematological findings in Kala‐azar, HIV, and Kalaazar co‐infected patients including T.lymphocytes differential count by both flowcytometry and double immunocytochemistry

Abstract

A descriptive study was conducted in two kalazar centers in Tabarak-Allah (Gedarif state) and El- Azaza (Singa state), Eastern Sudan. 160 VL patients out of 520 suspected VL cases were included. Diagnoses was made by direct microscopy using lymph node and/or B.M aspirates and confirmed serologicaly using DAT and/or Katex. There were 96(60%) males, 64(40%) females; mean of patient's ages was 14 years. Full blood count were done for all VL patients. Anemia was found in all, (mean of Hb: 8.5 g/dl), 80(50%) o them had a normocytic normochromic anemia, 16(10%) with microcytic hypochromic, 57(35.4%) with dimorphic anemia, and sickle cells anemia was found in 7 (4.6%) patients. 152 (95%) patients had a platelet count of < 140X103/ μl, 128(80%) patients with TWBCs < 3X 103/μl. The differential WBCs count was a follows: lymphocytes were > 40% in 112(70%) patients, neutrophils were < 45% in 89(55%) patients, and ESR was > 45 mm/h in 50/50 patients (100%). WBCs morphology was normal in 110(69%) patients, shift to left was found in 40(25%) patients, and shift to right was found in 6(10%) patients. Prominent Target cells were found in 54(34%) patients and nucleated RBCs in 24(15%) patients. The differential T cells count was done for 5 patients. CD4 T cells were < 250 / μl in all 5 patients. The mean of CD4 was 210 / μl, the mean of CD8 was 490 / μl, and CD4/CD8 ratio was 1/2.3. II All parasitologicaly and serologicaly confirmed VL patients were screened for HIV, 6 (3.5%) patients were co-infected, all of them were males, mean of patients ages was 25 years. Full blood counts were done for these patients. All patients were anemic (mean of Hb: 8.0 g/dl), 3(60%) patients with normocytic normochromic, 2(40%) patients with microcytic hypochromic anemia. Platelets were < 140X103, and TWBCs were < 3X 103/μl in all patients. The differential WBCs count was a follows: lymphocytes were > 40%, neutrophils were < 45%, and ESR > 45 mm/h were found in all patients. Also differential T cells count were done, CD4 T. cells were < 200/ μl in all patients, mean of CD4 was 180 / μl, mean of CD8 was 340 / μl, and CD4/CD8 ration was 1/1.9. A cross sectional study was done to determine the hematological parameters and T cell subsets count in 30 known seropositive HIV individual, during ART at VCT Centre (Om durman Teaching Hospital), Khartoum state. Anemia was found in 15(50%)patients (mean of Hb: 11g/dl), 12(80%) patients with normocytic normochromic, 3(20%) patients with microcytic hypochromic anemia. 4(13.3%) patients had platelets < 140X103, 5(16.7%) patients with TWBCs < 3X 103/μl, 10(33.3%) patients with TWBCs > 6X 103/μl. The differential WBCs count; lymphocytes were > 40% in 9(30%) patients, neutrophils were < 45% in 7(23.3%) patients, neutrophils were > 70% in 3(10%) patients. In differential T cells count, CD4 T. cells were < 200/ μl in 20(66.6%) patients, mean of CD4 was 255 / μl, mean of CD8 was 530 / μl, and CD4/CD8 ration was 1/2.1. We concluded that normocytic normochromic anemia, leucopenia, thrombocytopenia were the common hematological changes in VL and VL/HIV co-infected patients. Relative lymphocytosis and neutropenia were less common. In HIV seropositive individuals anemia were noted in have of patients. Luecocytosis, lymphocytosis, Eosinophilia and monocytosis were noted in third of patients. The differential T cells count indicated that, the cellular immunity was depressed among the VL, HIV, and VL / HIV co-infected patients; with decreased in CD4 number and reversed CD4/CD8 ratio. Comparison between Flowcytometry & double immunoenzymatic for CD4 counts were done for five HIV seropositive individual. Results indicated that there is a significant variation between the two methods (P <0.05). Sensitivity and specificity of double immunoenzymatic staining method were 100%.