دراسة جزيئية ومناعية لطفيلي اللشمانيا الجلدية Cutaneous Leishmania في المحافظات الوسطى والجنوبية

Molecular and Immunological study of Cutaneous Leishmania in the middle and southern provinces

The study includes distribution of the Cutaneous Leishmania in some governorates in Iraq. Specimens are included cases of outpatients in hospitals like: Al-Karamah, Al-Sadir Teaching Hospital in Baghdad, Al-Hilla Teaching Hospital, Al-Hussein Teaching Hospital and Ein Altamer General Hospitals in Kerbala ,Al-Sadir Teaching Hospital in Al-Najaf  , Al-Karamah and Al-Zahraa Teaching Hospitals in Wasit , Al-Diwanyia General Hospital . Al-Hussein General Hospital in Al-Nasiriya , Al-Smawa General Hospital, Al-Sadir Teaching Hospital and Al-Qurna General Hospital in Al-Basrah during October 2011  to march  2013

     Cutaneous Leishmania is consider a major problem that faces the patients because it causes deformities in the infection region .  The study includes 330 cases of skin ulcer in which 225 of them were microscopically positive. Parasitic growth was done using two types of cultural media NNN and RPMI-1640 .

The results have shown a considerable difference between male and female in which male was significantly exceeding 131  (58.22%) and female was 94(41.77%)  and 139 (61.77%) of outpatients from rural areas increased in numbers from outpatients of urban areas, and only (38.22%)from urban areas .

The number of ulcerations in body has exceeded infection ,that means more than one ulceration in male  83 (85%) and for female 41 (62%) , and concerning infection in three or four ulceration, it is approximately equal and normally distributed on face areas 99 and a percentage of (44%) then lower arms are  92 and a percentage of (40.80%) and the upper parts are 33 and a percentage of (14.66%). Also, one infection was recorded on the

The study shows that 46 patients in a percentage of (%20.44)  having  a dry infection;  whereas 179 patients (%79.55) having the wet type .

For accurate diagnosis of Leishmaniaa species distributed in Iraq, PCR Was used technique and diagnosed two types of Leishmania parasite that causes the illness. A band in a length of 560 bp in 186 samples related to Leishmania major and the band sample was 750 bp related to 39 Leishmania tropica.

Through studying the samples in Kerbala City, the number infected samples were 125 for the years of  2010-2011 the actuality is (73) infection and (52) infection from 2011-2012 in a ratio of (58.4 % and 41.6 %  accordingly) in different inhibited areas in the governorate  in which (51) (40.8%) was in Ein Al-Tamir Suburb ,  and 29 cases of  (23.2%)in Al-Husseinya Suburb , as well as 17 cases of (13.6%) in Al-Hur Suburb , 16 cases (12.8%) in the south quarters and 7 cases of (5.6%) in the north quarters and only 5 cases of (4%) in the city center .

The molecular diagnosis in Polymerase Chain Reaction for Kerbala governorate samples have shown 31 (24.8%) of L .tropica , 22 (17.6% )of  them were male and 9 of  (7.2%) of them were female . L. major was 94 of  (75.2%) in which 55 of (44%) were in male and 39  of (31.2% ) were in female .

     The immunological study for the patients serums  with L. major by ELISA technique has significantly shown raised values of IgG and IgM during infection in comparison with the control group , then it declines slowly after treatment in which IgG was in ratio (1811.1 ± 523.1 mg/dl) , and IgM was in ratio (166.7 ±23.6 mg/dl). Also, in  L. tropica – IgG was in ratio (1722.1 ± 524.0mg/dl) and IgM was in ratio (25.3±182.9mg/dl) in comparison with control specimens , and then the ratios have declined after treatment dosages with Pentostam drug.

      The cellular dynamics values have shown an abstract increase in which interferon- gamma (IFN-γ) in infected patients with cutaneous L. major were (5.5±113.2mg/dl) which declined after treatment abstractly to (3.0±6.05mg/dl) as well as for L tropica (6.5±88.2 mg/dl) which shows no significant abstract difference after treatment with control .

An increase appears in cytokine IL-10 that reached in L.major (9.8±215.0mg/dl) and after treatment was (5.1±9.02mg/dl) and in L. tropica was (8.8±115.0mg/dl) with no significant abstract difference after treatment with control specimen.

As a simple attempt to find a vaccine for Leishmania disease a Lipophosphoglycan was isolated and purified as known factor for promastigot stage. The vaccine injected for two groups of Bulb mice in a concentration of purified vaccine (L.majorand Ltropica) to study the immunological response by detecting the Lymphocyte Transformation assay ,the Delayed Type Hypersensitivity test and the PhagocytosisIndex  .

In lymphocyte transformation  the ratio of L. tropica was 6.5 %  and for L. major was 12.6 %  and there is significant difference by P˂ 0.05 in comparison with control specimen in which its ratio was 4.7%, and for Delayed Hypersensitivity test study the average foot thickness injected with vaccine and the other foot which injected with phenol-saline solution –only (1.21 ± 0.20ml) for the first group and (1.45 ± 0.10) for the second group ,and for the control group it has registered (0.0 ±0.445 ml) with a considerable difference of P˂ 0.05 . By calculating the ratio of Phagocytosis Index cells, it was 17% and 27.6% for the first and second groups while for the control group it has been reaching 9.10% . Therefore, we can get advantage from these values as indicators for the rule of vaccine in the immunological response and triggering of vaccine idea for this disease .