Somaya El Deeb

Effect of hydrocortisone on immune system of the lizard, Chalcides ocellatus. III: Effect on cellular and humoral immune responses

Administration of a single injection of 1.0 mg/g body weight hydrocortisone acetate (HC) to adult lizards, Chalcides ocellatus in summer elicited a high and lasting rise in serum corticosterone and cortisol, peak levels being, however, in the physiologic ranges of 10 micrograms % (2 X 10(-7) M) and 40 micrograms % (1 X 10(-6) M), respectively. Elevation of serum corticosteroid (CS) concentrations by exogenous HC impaired the proliferative capacity of spleen cells in mixed leukocyte reaction (MLR) and significantly delayed skin allograft rejection. In vivo HC also abrogated the primary in vivo and in vitro immune responses of lizards to rat erythrocytes (RRBC). Reactivity to allogeneic cells in MLR, and antibody production against RRBC were, however, recovered when serum CS levels resumed normal, basal values i.e. at three weeks post-HC injection. These data indicate that rise in circulating CS induces severe immunosuppression in lizards.

Ontogeny of con a responsiveness and mixed leucocyte reactivity in the lizard, Chalcidesocellatus

The immune system of phylogenetically key animals will contribute significantly to our understanding of the evolution of immune response in higher vertebrates. Reptiles, being evolutionary precursors of both birds and mammals, represent a pivotal group and thus a study of their immune system is of special significance. Here we described the emergence of T-cell immune capability in the viviparous lizard (Chalcides ocellatus) throughout embryonic development (stages 36-41 of Zada and El Deeb, 1984) and in newborns. The response of embryonic thymocytes (5 X 10(5) cells/ml) to Con A (5 micrograms/ml in culture) was first detected at stages 36-37, increased gradually during successive stages and then declined at birth to yield low responses in newborn lizards. In addition, embryonic thymocytes cultured in two-way MLR, using several combination sets, were significantly responsive at all stages. Our results reveal a degree of immunological T-cell maturation during reptitilian embryonic life which is similar to results in amphibians and mammals, but not clear with respect to fish and birds where comparative information still somewhat limited.

Ontogeny of hemopoietic and lymphopoietic tissues in the lizard Chalcides ocellatus (Reptilia, Sauna, Scincidae)

The first and major blood‐forming organ to develop in the viviparous lizard Chalcides ocellatus is the yolk sac, which exhibits prominent erythropoietic activity from as early as stage 21 through birth (stage 41). Myeloid cells and megakaryocytes are produced in the yolk sac from stage 23 onward. During lizard embryogenesis hemopoietic activity is also observed in spleen and bone marrow but in neither kidney nor liver. Cells capable of giving rise to lymphocytes both in vivo and in vitro are first found in the thymus at stage 35. Active lymphopolesis in thymus and spleen begins at stages 36 and 39, respectively. In contrast, the gut‐associated lymphoid aggregates are not evident before birth.

The development of lymphocytes with T- or B- membrane determinants in the lizard embryo

To study the ontogeny of lymphocytes in the lizard, Chalcides ocellatus antisera were raised in rabbits against thymocytes collected from adult lizards (AATS) or from embryos of stages 40 and 41 (AETS), and against serum immunoglobulins (AGGS). AATS recognized in adult lizards a surface membrane antigenic system (TA) specific to thymocytes and thymus-derived (T-) cells in spleen, and a distinct antigenic entity (Tt) found exclusively on intrathymic lymphocytes. After exhaustive absorption with adult thymocytes, AETS defined a further antigen (TE). On the other hand, AGGS combined with the immunoglobulin (Ig) determinants present in the cytoplasm and surface membrane of lizard presumptive B lymphocytes. Immunofluorescence assays during the development of the T and B lineage showed that TA+ and Tt+ thymocytes increased from 35% at stage 37 to 96% at stage 41, and were exported precociously to the embryonic splenic environment. Embryonic thymocytes were shown to totally lack surface Ig, whereas 40-50% of splenocytes carried cytoplasmic and/or surface membrane Ig determinants. The data revealed further the presence of a subset of splenocytes bearing the specific markers of both T and B cells.

Characterization of diverse hemolymph lectins in the cotton caterpillar Spodoptera littoralis

Abstract 1. 1. Hemolymph lectins (agglutinins) of the cotton caterpillar Spodoptera littoralis were analyzed by agglutination, cross-absorption and carbohydrate-hemagglutination inhibition using several vertebrate erythrocytes. 2. 2. Lectins were found to interact, with all tested erythrocytes, by binding to carbohydrate moieties but showing no definite specificity. 3. 3. Disulphide bonds were probably absent as 2-ME treatment was ineffective. 4. 4. By cross-absorption studies, we have proposed that the hemolymph contains multiple lectins.

Effect of artemether on cytokine profile and egg induced pathology in murine schistosomiasis mansoni.

Artemether (ART), the methylated derivative of artemisinin, is an efficacious antimalarial drug that also displays antischistosomal properties. This study was designed to evaluate the immunomodulatory action of a single intramuscular dose (50 mg/kg body weight) of ART in comparison with PZQ treatment (42 days PI). ART administration was 7, 14, 21 and 45 days PI. ART effect was studied parasitologically, histopathologically and immunologically. It was found that maximum effect was reached when ART treatment interfered with 14 or 21 days old schistosomula. ART treatment 14 or 21 days PI was associated with shift from Th2 to Th1 predominancy (decrease in IL-4 and upgrading of serum IFN-γ levels). In conclusion, ART is a promising drug in control of schistosomiasis mansoni due to its reductive effect on worm burden and its role in improvement of hepatic granulomatous lesions.

Immunodiagnosis of fascioliasis using sandwich enzyme-linked immunosorbent assay for detection of Fasciola gigantica paramyosin antigen.

Background: Many immunological techniques have been developed over years using the different Fasciola antigens for diagnosis of parasitic infection and to replace the parasitological techniques, which are time consuming and usually lack sensitivity and reproducibility. Materials and Methods: In this study, Fasciola gigantica paramyosin (Pmy) antigen was early detected in cattle sera using sandwich enzyme-linked immunosorbent assay (ELISA), to evaluate the Pmy antigen performance in diagnosis. This work was conducted on 135 cattle blood samples, which were classified according to parasitological investigation into, healthy control (30), fascioliasis (75), and other parasites (30) groups. Results: The sensitivity of Sandwich ELISA was 97.33%, and the specificity was 95%, in comparison with parasitological examination, which recorded 66.66% sensitivity and 100% specificity, respectively. Conclusions: It was clear that the native F. gigantica Pmy is considered as a powerful antigen in early immunodiagnosis of fascioliasis, using a highly sensitive and specific sandwich ELISA technique.

Role of relevant immune-modulators and cytokines in hepatocellular carcinoma and premalignant hepatic lesions

AIM To assess the levels of different immune modulators in patients with hepatocellular carcinoma (HCC), in relation to other hepatic diseases. METHODS Eighty-eight patients were included in the current study and represented patients with HCC (20), liver cirrhosis (28) and chronic hepatitis (CH; 25), and normal controls (NC; 15). Peripheral blood was isolated for immunophenotyping of active myeloid dendritic cells (mDCs; CD1c and CD40), mature inactive myeloid cells (CD1c and HLA), active plasmacytoid cells (pDCs; CD303 and CD40), mature inactive pDCs (CD30 and HLA), active natural killer (NK) cells (CD56 and CD161), active NK cells (CD56 and CD314) and inactive NK cells (CD56 and CD158) was done by flow cytometry. Serum levels of interleukin (IL)-2, IL-10, IL-12, IL-1β, interferon (IFN)-α, IFN-γ and tumor necrosis factor (TNF)-αR2 were assessed by ELISA. RESULTS Active mDCs (CD1C+/CD40+) and inactive mDCs (CD1c+/HLA+) were significantly decreased in HCC patients in relation to NC (P < 0.001). CD40+ expression on active pDCs was decreased in HCC patients (P < 0.001), and its level was not significantly changed among other groups. Inactive pDCs (CD303+/HLA+), inactive NKs (CD56+/CD158+) and active NKs (CD56+/CD161+) were not statistically changed among the four groups studied; however, the latter was increased in CH (P < 0.05). NKG2D was statistically decreased in HCC, CH and cirrhosis (P < 0.001), and it was not expressed in 63% (12/20) of HCC patients. There was significant decrease of IL-2, IFN-α and IFN-γ (P < 0.001), and a significant increase in IL-10, IL-1β, and TNF-αR2 (P <0.01, P < 0.001 and P < 0.001; respectively) in HCC patients. There was inverted correlation between IL-12 and IL-1β in HCC (r = -0.565, P < 0.01), with a strong correlation between pDCs (CD303+/CD40+) and NKs (CD56+/CD161+; r = 0.512, P < 0.05) as well as inactive mDCs (CD1c+/HLA+) and inactive NK cells (CD56+/CD158+; r = 0.945, P < 0.001). CONCLUSION NKG2D, CD40, IL-2 and IL-10 are important modulators in the development and progression of HCC.