Flow cytometric analysis of leishmania reactive CD4[+]/CD8[+] lymphocyte proliferation in cutaneous leishmaniasis

Determination of the division histoty of T cells in vitro is helpful in the study of effector mechanisms against infections. Technique described here uses the intracellular fluorescent label carboxyfluorescein diacetate succinimidyl ester [CFSE] to monitor the proliferation. In a cross sectional study, blood samples were collected from 7 volunteers with history of cutaneous leishmaniasis [CL] and one healthy control from endemic areas in Isfahan province who referred to the Center for Research and Training in Skin Diseases and Leprosy [CRTSDL], then CD4[+]/CD8[+] lymphocytes and CD14[+] monocytes were isolated from peripheral blood mononuclear cells [PBMC] using mAbs and magnetic nanoparticles. CFSE labeled CD4[+] or CD8[+] lymphocytes cultured with autologous monocytes in the presence of PHA, SLA, live Leishmania major or as control without stimulation. Cells were harvested after 7 days and were analyzed using flow cytometry. Five consecutive divisions were monitored separately. Stimulation of CD4[+] or CD8[+] lymphocytes from CL subjects with SLA showed a significant difference in proliferation comparing with unstimulated cells [P< 0.05]. The significant difference in the percentages of CD4[+] cells stimulated with SLA was revealed at different divisions for each subject. In CD8[+] lymphocyte, significant stronger stimulation of SLA was evident later in the proliferation process. The mean number of divisions in both CD4[+]/CD8[+] lymphocytes stimulated with SLA was significantly greater than when stimulated with live L. major [P=0.007 / P=0.012, respectively]. The percentage of divided cells might be calculated separately in each division. The cells remained active following CFSE staining and there is possibility of functional analysis simultaneously

Intestinal parasitic infections in renal transplant recipients

Organ transplant recipients can experience serious diseases from infections due to emerging and reemerging parasitic infections. This study was carried out to evaluate the prevalence of intestinal parasites among renal transplant recipients of Iran. This cross-sectional study was conducted from June 2003 to August 2004 on renal transplant recipients in Iran. A total of 706 fecal samples obtained from randomly selected population originated from all over Iran. Patient's information was recorded in a questionnaire before sampling. A sample of stool was taken from each person. Direct wet smear examination, formalin-ether concentration, Ziehl-neelsen staining, and agar plate culture were done for each sample. Totally 32 patients [4.5%] were positive for parasitic infections. In searching for emerging parasitic infections, the most prevalent parasites were found to be Blastocystis hominis, Giardia lamblia and Entamoeba coli, respectively. The merely ova which were seen were related to Hymenolepis nana. With investigation of healthy control, no significant difference was found between transplanted and normal population. The population showed controlled rate of intestinal infections probably due to regular awareness concerning risks of opportunistic infections; albeit regular surveillance through routine examination of stool samples for parasites seems considerably advantages the transplant recipient patients

Serological study of toxoplasma gondii infection using IFA method in renal transplant recipients

Toxoplasmosis is a wide distributed opportunistic infection caused by Toxoplasma gondii. This was a cross-sectional study of T. gondii antibody titer, which was conducted from June 2003 to August 2004 on renal transplant recipients in Iran. A total of 551 serum samples were obtained from randomly selected population referred from different areas all over the country to Shafa Central Clinic in Tehran. Patient's information was recorded in a questionnaire before sampling. Two samples of finger-prick blood were collected from each person and antibody titer against Toxoplasma was assessed by Indirect Fluorescence Antibody [IFA] technique on serum samples. Totally 39 cases [7.1%] of samples were positive for antibody by the titer of 1: 20 and higher. On investigation of risk factors, no significant difference was found between consumption of under-cooked meat, close contact with animals, and the source of drinking water and seropositivity rate of toxoplasmosis. The relatively low seroprevalence rate of Toxoplasma infection shows the successful approaches to awareness of transplant recipients about the potential risks of acquisition of infectious diseases due to regular administration of suppressive drugs. However, the regular surveillance through serological screening of Toxoplasma antibody in kidney transplant recipients is advisable