Post challenging serum cytokine profile (Th1 & Th2) in the vaccinated mice (Balb/C) with a new formulation of Leishmania major antigen.

OBJECTIVE: The aim of this study was to carry out experiments further to our previous new formulation to modify the Leishmania major antigen that had satisfactory results previously. METHODS: In this study we made a preliminary, new vaccine with the same methodology and selected two injection doses (100&200 µg/o.1 mL), three injection Groups: Leishmania plus BCG (LB), Leishmania plus new adjuvant (Teucrium Polium) [LT], Leishmania plus BCG and Teucrium Polium (LBT), and one susceptible mouse Group (Balb/c) and measure two types of cytokines: Th1 (IFN-γ, IL-12) and Th2 (IL-4, IL-10) We prepared crude antigen combinations by five different methods using antigens from L. major parasites. Phase I was done in the animal model. In our study, Leishmania antigen was examined both with BCG and the new adjuvant (TP) in three Groups in two injection doses (100.200 µg/1 mL) and Balb/c mice. RESULTS: Our results showed that in three injection Groups (LB, LT and LBT) that received each or both BCG and TP as adjutant with injection doses of 100 and 200 µg/1 mL with two booster doses: the LBT Group had the lowest IFNγ and highest IL-12 value, LT and LB Groups have equal IL-12, but LB have more IFNγ and IL-10 but less than IL-4 in the LT Group. CONCLUSION: In this study, the LBT Group has statistical differences regarding IL-12 and IL-10 from the other Groups.

Antioxidants and proinflamatory cytokines in the sera of patients with cutaneous leishmaniasis.

BACKGROUND: Leishmania is a significant health problem in many parts of the world. Tumor necrosis factor (TNF) plays an essential role in Leishmania major infections. OBJECTIVE: To study the pro-inflammatory cytokines and antioxidants in four groups of cutaneous leishmaniasis patients. METHODS: 39 patients were divided into four groups of: 1) active (acute phase of treatment); 2) non-healing (received treatment for almost two years without recovery); 3) healing (recovered upon treatment); and 4) healed (previously received treatment and achieved complete remission) patients. Serum levels of pro-inflammatory cytokines (IL-1B, TNF-α, IL-6) and serum antioxidant levels were measured by ELISA and FRAP assays, respectively. RESULTS: While serum antioxidant levels were elevated in the non-healing group, there was no difference among other groups of patients and healthy controls in this regard. Interleukin-1ß showed the highest level in the non-healing group followed by the other groups of patients. The mean serum IL-6 level was highest in the non-healing group, but showed no significant change in the other groups. TNF-α and IL-1ß levels were non-significantly elevated in the sera of active and non-healing patients. CONCLUSION: Pro-inflammatory cytokines IL-1ß, TNF-α, IL-6 maybe related to the progression of leishmaniasis. Serum antioxidant levels maybe correlated with patient response to drug treatment.

Evaluation of the effects of a new formulation of Leishmania major antigen in Balb/C and conventional white laboratory mice.

BACKGROUND/PURPOSE: Leishmaniasis is currently a threat in 80 countries around the world, and cutaneous Leishmaniasis accounts for more than 5% of new cases. It is a problematic disease in Iran, and preparing a protective vaccine has been a major goal of medical investigations. The objective of this study was to compare the protective effects of a cocktail vaccine candidate encoding various Leishmania major antigens in highly susceptible (Balb/C, or type I mice) and resistant (laboratory small white, or type II) mice. METHODS: A new antigen formulation was evaluated in type I and II mice. Leishmania major promastigotes was cultured and harvested at different growth stages, and a cocktail made from the harvested organisms. The preparations were tested for sterility and contamination by endotoxin. Five different methods were utilized to produce a crude antigen preparation. The protein levels of the antigen preparations were measured using the Lowry method, and the antigens were intradermally injected using different protocols for type I and II mice. After 38 days, all mice were euthanized with diethyl ether, and spleens were removed. Histological sections were prepared, stained with hematoxylin and eosin and changes to the splenic white pulp (SWP) were studied microscopically. RESULTS: Compared with the control groups, there was a drastic change in white pulp structure. The size increases of the SWP were dependent on the injection group and mouse strain. There was a remarkable expansion of lymphoid follicles in the treated groups in both mice strains. CONCLUSION: The new antigen formulation was able to stimulate and expand the lymphoid constituents of spleen tissue. The SWP is where immune responses and antibodies are produced. Therefore, the effect of the antigen preparations on secondary immune responses, adaptive immunity, and antibody production is important in determining the susceptibility of mice to cutaneous Leishmaniasis and the induction of immunity.

Neutrophil function (innate immunity) during Ramadan.

BACKGROUND: Fasting during the month of Ramadan is one of the essential religious practices of Muslims. The aim of this study was to evaluate opsonisation, phagocytosis, and nitroblue tetrazolium (NBT) reduction by white blood cells in normal, healthy, male subjects under non-fasting (before Ramadan) and fasting (after Ramadan) conditions. METHODS: In this study, 13 Muslim men, aged 28-54 years, whose health was confirmed by health application forms, gave blood samples one week before the beginning of the holy month of Ramadan and during the last week of Ramadan. Blood samples were tested for neutrophil phagocytosis, serum opsonisation power, and NBT reduction. RESULTS: Despite a decline in the neutrophil phagocytic index and serum opsonisation index, the percentage of neutrophils participating in phagocytosis increased with fasting. In addition, there was an increase in the percentage of neutrophils demonstrating NBT reduction. Although there was a decrease in opsonisation of the serum, the increased percentage of opsonisation compensated for this defect. CONCLUSION: This study demonstrates the beneficial effect of fasting during Ramadan on neutrophil phagocytic function.

Safety and toxicity of a new formulated Leishmania major preliminary vaccine in animal model Balb/c and small white conventional laboratory mice.

Leishmania parasites cause a spectrum of diseases that afflict the populations of 88 countries around the world and all attempts to control leishmaniasis have failed. It seems that preparing a vaccine may be the final solution. The aim of this study was to determine various Leishmania (L.) major antigens vaccine candidates and effects of the vaccine on delayed type hypersensitivity (DTH). Many different methods of vaccine preparation plus or without adjuvant were used. We prepared crude antigen combinations by five different methods using antigens from L. major parasites. Phase I was done in animals. The immunogenic effect was evaluated with the delayed type hypersensitivity (DTH) reaction with five different doses, including 100, 200, 300, 400, 500 microg/ml of total protein + BCG in Balb/c and conventional laboratory white mice (out breed). Our results showed that the cocktail antigen was highly specific. No injection of BCG solvent or saline treated controls showed significant results. Taken together, the effect of cellular immune response to the cocktail vaccine induced a significant effect against cutaneous leishmaniasis in the experimental model of vaccine with L. major.