Iranian journal of immunology
Volume:10 Issue: 2, Spring 2013

Astrocytes, which comprise ~90% of overall brain mass, are involved in brain immunity. These cells represent the non-professional class of CNS-resident APCs and may promote or inhibit CNS inflammation depending on the cytokines they secrete. IL-10 family of cytokines and their receptors, IL-20R1 and IL-20R2, may have a role in shifting astrocytes to a neuroprotective or neurodegenerative function. To address the expression of IL-20R1 and IL-20R2 cytokine receptors in astrocytes andbrain cortex of C57BL/6 mice. We investigated the expression of IL-20R1 and IL-20R2 in C57BL/6 mice astroglial cells and brain cortex in response to lipopolysaccharide (LPS), using reverse-transcription polymerase chain reaction (RTPCR) method. Astrocytes were able to express IL-20R1 and IL-20R2 mRNA not only in response to LPS stimulation but also in the absence of LPS. Furthermore, we found the expression of IL-20R1 and IL-20R2 mRNA in the cortex of adult C57BL/6 mice. IL-20R1 and IL-20R2 are constitutively express in the brain. Since most neuropathological processes involve astrocytes and inflammatory cytokines, these findings have important implications for future therapeutic strategies. Abd Nikfarjam B, et al. Iran J Immunol.

Noradrenaline (NA), the principal neurotransmitter released from sympathetic nerve terminals, influences T-cell maturation, not only directly in developing T cells, but also indirectly, by acting on the thymic nonlymphoid cells. In vitro and in vivo studies have demonstrated the anti-proliferative, anti-migratory, antiangiogenic and cytotoxic properties of propranolol, β-AR blocker, against various cancers. To evaluate the effect of propranolol on efficacy of HSP-70 rich lysate vaccine in immunotherapy of fibrosarcoma. Mouse fibrosarcoma WEHI-164 cells were used to immunize tumor-bearing mice with or without propranolol and HSP-70. Splenocytes proliferation, cytotoxicity activity of the splenocytes, naturally occurring CD4+ CD25high T-reg cells and IFN-γ and IL-4 secretion as well as tumor size, were assessed to describe the anti-tumor immune response. A significant increase in the level of IFN-γ in the mice vaccinated with WEHI-164 cells enriched with HSP-70 and co-treated with propranolol was observed compared to controls. However, HSP enrichment or propranolol treatment alone did not enhance the immune response as measured by the level of IFN-γ. Likewise, a decrease in tumor growth in the test group (p<0.01) and a significant increase in CTL activity (p<0.05) was observed. HSP enriched vaccine shows anti-tumor activity, probably due to the modulation of immune responses.

Inflammatory bowel disease, an autoimmune disease, has two clinical manifestations including Crohn’s disease and ulcerative colitis (UC). IL-17 has been the target of intensive research in autoimmune diseases. The influence of Toll like receptor 4 (TLR-4) gene polymorphisms on IL-17 production has also been revealed in UC patients and tissue inflammation in mice. To investigate the association between the TLR-4 gene polymorphisms, Asp299Gly and Thr399Ile and IL-17 serum levels with ulcerative colitis. Additionally, we aimed to study modulation effects of forenamed gene polymorphisms on IL-17 serum levels in UC patients and controls. A total of 256 healthy controls and 85 UC patients enrolled in our study. DNA was extracted and PCR-RFLP technique was employed to determine Asp299Gly and Thr399Ile polymorphisms in TLR-4 gene and IL-17 serum levels were measured by ELISA method. There was no significant difference between the frequency of Asp299Gly A>G and Thr399Ile C>T in UC patients and controls. While IL-17 serum levels in UC patients were significantly higher than controls (p=0.003), no significant difference in IL-17 levels between different genotypes existed. Additionally, a significant inverse relationship was observed between hemoglobin level and IL-17 serum levels in UC patients (p=0.039). Increased IL-17 serum levels in our UC patients might be explained through the synergistic activity of IL-17/IL-23 axis and pro-inflammatory cytokines, causing severe clinical outcome in patients with IBD. The prolonged excretion of blood in stool driven by inflammatory process which causes iron metabolism disorder and anemia may elucidate the inverse correlation between hemoglobin and IL-17 serum levels in UC patients. Lack of association between the TLR-4 gene polymorphisms and UC in our study was consistent with the results from other Caucasian populations.

Although there is convincing data in support of the effectiveness of hyperthermia in tumor therapy, the molecular mechanisms underlying the clinical effects of hyperthermia are still poorly understood. To investigate natural killer (NK) cell cytotoxicity against heat-treated SW-872 and HeLa tumor cell lines. NKG2D ligands and HLA class I transcription were examined using quantitative real-time PCR in treated tumor cell lines at 0, 2, 4, 6 and 12 h following thermal treatment at 39C and 42C for 1 h. The expression of MICA/B, ULBP1 and ULBP2 were also determined by flow cytometry. NK92-MI cytotoxic activity against heat-treated target cell lines was assessed by LDH release as well as annexin-V and 7- AAD assays. Our results showed that heat treatment at 39C improved the cytolytic activity of NK cells against SW-872 cells without increasing NKG2D ligand concentration or decreasing HLA class I levels. The observed increase in the cytotoxicity of NK cells against SW-872 cells after hyperthermia does not coincide with changes in MICA/B, ULBP1 and ULBP2 ligands of NKG2, however, the expression of other ligands in target cells may have made the cells susceptible to the cytotoxic effect of NK cells.

Absent in Melanoma 2 (AIM2) is an intracellular microbial dsDNA sensor which plays an important role in production of proinflammatory cytokines through Apoptosis associated Speck-like protein containing a Caspase activation and recruitment domain (ASC) and Caspase-1. Micro-RNAs (miRNAs) play important roles in regulation of immune related genes. However, there is little information regarding the effects of miRNAs on the AIM2 and ASC expression. To determine the mRNA levels of AIM2 and ASC in Jurkat cell line following introducing miRNA-143 (miR-143). MiR-143, a scrambled sequence and PBS were introduced separately, to the Jurkat cell lines and the mRNA levels of AIM2 and ASC were examined in parallel with beta-actin and GAPDH (as housekeeping genes) using Real- Time PCR technique. The mRNA levels of AIM2 and ASC were significantly increased in the miR-143 transfected Jurkat cells when compared to the scrambled sequence or PBS treated cells. miR-143 can lead to increased expression of AIM2 and ASC mRNAs. Considering the significance of AIM2 and ASC in DNA sensing and inflammosome formation, it can be considered as a therapeutic agent for the treatment of chronic infectious diseases, especially viral infections.

Recombinant vaccine technology is one of the most developed means in controlling infectious diseases. However, an effective vaccine against Shigella is still missing. To evaluate recombinant IpaC protein of Shigella as a vaccine candidate. In this study we cloned ipaC gene into an expression vector in prokaryotic system. The protein expression was evaluated by SDS-PAGE and Western- Blotting analysis. The recombinant protein was purified using Ni–NTA affinity chromatography. Guinea pigs were immunized with the recombinant protein and the level of immunogenicity was examined by ELISA and Western blotting of IpaC. Challenge test was done through the intraoculary injection of Shigella dysenteriae (6×108 CFU/eye) and after 48 hours was scored for keratoconjunctivitis. The results showed a remarkable level of immunogenicity in terms of antibody response and protection against keratoconjunctivitis in tested animals. The recombinant IpaC protein provided a protective system against Shigella dysenteriae type I during the challenge test. The results showed the potential of using recombinant IpaC in preparation of vaccine in perspective studies.

Iron Deficiency Anemia (IDA) has been controversially linked to IL-4 production in previous studies. A predominant Th1 response leads to resistance against recurrent vulvovaginal candidiasis (RVVC), whereas a Th2 response exacerbates the disease. To investigate the possible effect of iron deficiency on the host’s susceptibility to RVVC as a result of the Th1/Th2 cytokine polarization. We conducted a case-control study of 92 women in 4 groups based on strict inclusion and exclusion criteria: RVVC+IDA+ group consisted of 23 women with RVVC and IDA; RVVC+ IDA- group consisted of 23 women with RVVC without IDA; RVVC-IDA+ group consisted of 23 women without RVVC and with IDA and RVVC- IDA- group consisted of 23 healthy women. The iron parameters and key cytokines (IFN-γ, IL-10, IL- 12, IL-4) were measured in blood samples. Comparison of IL-4 production between RVVC+ IDA+ (12.2 ± 1.3 pg/ml) and RVVC+ IDA- (2.4 ± 4.0 pg/ml) groups (p=0.044), between RVVC- IDA+ (14.6 ± 1.7 pg/ml) and RVVC- IDA- (1.28 ± 3.6 pg/ml) groups (p=0.006), between RVVC- IDA+ (14.6 ± 1.7 pg/ml) and RVVC+ IDA-) 2.4 ± 4.0 pg/ml) groups (p=0.009) and also between RVVC+ IDA+ and RVVC- IDA- (1.28 ± 3.6 pg/ml) groups (p=0.03) showed significant differences. We found a significant positive correlation between IL-4 and total iron binding capacity (TIBC, p=0.046) and between serum IL-10 and Hb levels (p=0.041) in the RVVC+ IDA- group. There was also a significant negative correlation between serum IL-4 and levels of serum iron (SI, p=0.041) in the RVVC- IDA- group. It seems that IDA determines the balance between and the intensity of Th1 and Th2 arms of the immune response and leads to a deviation toward Th2 response which could contribute to recurrence of candidiasis.