Association of KIR3DS1+HLA-B Bw4[Ile80] combination with susceptibility to tuberculosis in Lur population of Iran

Natural killer [NK] cells are the effector cells of innate immunity that respond to infection and tumor. Interactions between killer cell immunoglobulin like receptors [KIR] and human leukocyte antigen [HLA] class I molecules regulate NK cells responses to eliminate infected and transformed cells. To investigate the impact of KIR genes, HLA ligand genes, and KIR-HLA combinations on susceptibility to tuberculosis [TB] in Lur population of Iran. The genomic DNA of 50 patients with TB from Lorestan province of Iran was genotyped for sixteen KIR genes and their five major HLA class I ligands were determined by a polymerase chain reaction-sequence-specific primers [PCR-SSP] assay. The results were compared with those of 200 healthy unrelated Iranian individuals. In Lur population of Iran, a significant decrease in frequency of KIR3DS1 was found in TB patients compared to control group [24% vs. 44.5%, OR=0.394, CI=0.194-0.798, p=0.013]. Also, among the three activating genes that may use HLA class I molecules as their ligands, a significant decrease was shown in frequency of KIR3DS1 with HLA-B Bw4[Ile80] ligand in TB patients compared to control group [4% vs. 23%, OR=0.14, CI=0.033-0.596, p=0.004]. These findings imply a genetic imbalance between activating and inhibitory KIR genes and KIR-HLA combinations in Lur TB patients. Low level of activating KIR3DS1 and its combination with HLA-B Bw4[Ile80] ligand might have an influence on the susceptibility to TB in Lur population of Iran

Study of KIR expression and HLA ligands in cd56+ lymphocytes of drug resistant tuberculosis patients

Analysis of receptor-ligand interactions in the context of diseases necessitates to understand how HLA-KIR genotypes function in diseases. Although CD56+ lymphocytes are derived from multiple lineages, they share a functional association with immunosurviellance and antimicrobial responses. The present study aimed to determine whether KIR phenotype in CD56 lymphocytes and corresponding HLA-class 1 ligands are associated with multidrug resistance tuberculosis [MDR-TB]. We compared the frequencies of HLA-C and HLA-BW4 genes, the expression of KIRs 2DL1/2DS1, 2DL2/2DL3, 3DL1, and 2DS4 and the combinations of HLA/KIR in 32 Nifamycin and Isoniazid-resistant TB with those in 68 drug non resistant [NR] sputum smear positive pulmonary TB patients. PCR-SSP and flow cytometry were performed for HLA and KIRs typing, respectively. We showed no significant differences between inhibitory or activating KIRs as well as HLA ligands in MDR TB patients compared with NR-TB. The combinations of inhibitory KIR-HLA ligands in MDR-TB were much more prevalent, but not statistically significant than in NR patients [p=0.07]. The frequency of MDR patients with all HLA-C and HLABW4 ligands was higher than NR-TB [p<0.009]. Conversely, the percentage of MDR patients having only one kind of HLA gene was significantly lower than NR-TB [p<0.01]. We conclude that the expression of inhibitory KIRs with corresponding HLA ligands genes, and/or co-existence of three HLA class 1 ligands for inhibitory KIRs may be associated with drug resistance in pulmonary tuberculosis

Co-administration of Chenopodium album allergens and CpG oligodeoxy- nucleotides effects on peripheral blood mononuclear cells of patients with allergic rhinitis treated with intranasal corticosteroids and antihistamines

Allergic Rhinitis [AR] is one of the most common chronic diseases in the developed countries. This study was performed to investigate the effect of CpG-ODN in alteration of T-helper [Th]l/Th2 balance of patients with AR treated with intranasal corticosteroids [INCs] and antihistamines. Peripheral blood mononuclear cells [PBMCs] of 20 patients with AR were isolated before and after 45 days therapy. Cytokine production [IL-4, IL-10, IL-13, IFN-gamma] and specific Ch.a IgE in response to CpG co-administration of natural chenopodium album [CpG/Ch.a] or recombinant Ch.a [CpG/rCh.a] allergen were investigated in supernatants.of cultured PBMCs using ELISA Intracellular IL-10 was also assessed in CD4[+] cells using flow cytometry. Significant increase in production of IFN-y and IL-10 and decrease in production of IL-4 were found in supernatants of cultured PBMCs activated with CPG/ch.a and CPG/rch.a. of both CpG/Ch.a and CpG/rCh.a compared to allergens alone, before and after therapy. After therapy, IFN-gamma production with CpG/Ch.a was significantly increased in comparison with before [237 vs. 44 pg/ml, p=0.001]. IFN-gamma and IL-10 production with CpG/rCh.a was significantly increased after therapy compared to before [407.6 vs. 109 pg/ml, p=0.0l for IFN- gamma; 171.7 vs. 52.6 pg/ml, p=0.008 for IL-10], whilst IL-4 was significantly decreased [2.1 vs. 5.8 pg/ml, p=0.02]. Intracellular IL-10 expression was also significantly increased in response to either CpG/Ch.a or CpG/rCh.a that showed intracellular assay could be more sensitive than ELISA. Also, treatment with intranasal corticosteroids and antihistamines could enhance this CpG effect, in vitro

Inhibitory killer cell immunoglobulin-like KIR3DL1 in combination with HLA-B Bw4[iso] protect against ankylosing spondylitis

The HLA class I molecules serve as ligands for both T cell receptors and killer cell immunoglobulin-like receptors [KIRs]. We investigated the HLA-C and HLA-Bw4 alleles as well as KIRs expression on CD56 positive lymphocytes to evaluate whether these genes and molecules could influence Ankylosing spondylitis [AS] susceptibility, alone or in combination. We typed 40 AS patients and 40 normal controls for HLA-C asn[80] [group 1] and HLA-C lys[80] [group 2], HLA-B Bw4[thero], HLA-B Bw4[iso] and HLA-A Bw4 alleles by PCR-SSP method. We also assessed the expression of KIR2DL1/2DS1, KIR2DL2/2DL3, KIR3DLI and KIR2DS4 by flow cytometry. The Pearson chi-square or Fisher exact test was performed for statistical analysis. The frequency of HLA-B Bw4[iso] but not HLA-B Bw4[thero] and HLA-A Bw4, ligand for the inhibitory KIR3DL1, was significant reduced in AS patients as compared with controls [p<0.01]. No significant differences were observed in gene carrier frequencies of HLA-C group 1 and 2 between AS and controls. Although no differences were found in the expression of KIR receptors between AS and normal was reduced in patients with AS compared to healthy controls [p<0.009]. We conclude that HLA-B Bw4[iso], the ligand of inhibitory KIR3DL1, with and without the expression of KIR3DL1 might be involved in protection against AS. Our results suggest that besides the HLA and KIR genotype, expression levels of KIRs may be involved in the pathogenesis of AS disease

Elevation of CD56[bright]CD16[-] lymphocytes in MDR pulmonary tuberculosis

Protective immune responses induced in the majority of people infected with Mycobacterium tuberculosis enable them to control TB infection. The aim of this study was to investigate CD56 and CD16 positive peripheral blood mononuclear cells [PBMCs] and leukocyte subsets from multi-drug resistant pulmonary tuberculosis [MDR-TB], and compare them with nonresistant [NR] TB patients and healthy controls. 13 MDR-tuberculosis patients, 20 NR-TB individuals and 40 healthy subjects were included. Peripheral blood mononuclear cells were double stained with fluorochrome conjugated antibodies against CD56 and CD16 cell surface markers. The phenotype of positive cells was then analyzed by flow cytometry and the percentages of CD56[+] CD16[+], CD56[-] CD16[+], CD56[dim]CD16[ +/- ], and CD56[bright]CD16[ +/- ] subsets were calculated. There was a significant decline in the percentage of CD56[]+CD16[+] lymphocytes in both MDR and NR-TB patients compared with healthy controls. We also observed lower proportions of CD56[dim]/[bright]CD16[+] in addition to higher percentages of CD56dim/brightCD16-subsets in all TB patients [p

Immunophenotype of peripheral blood lymphocytes following thermal injury in patients

Severe immunosuppression occurs after large thermal burn and probably contributes substantially to patient morbidity and mortality.In this study we investigated the range and distribution of T-lymphocyte. Subsets CD3[+] [T cells] CD4[+] [T helper/inducer cell,.th], CD8[+] [T suppressor /Cytotoxic cells, TS/C], CD3[+] CD4VCD3[+]CD8[+] ratio, CD19[+] [B cells] and CD16[+] [NK cells] in patiens following thermal injury. Forty male, aging 18-60 years with major thermal injury were studied. The total body surface area of the burn injury, ranged from 30 to >70%. Whole blood samples were collected at three and seven days postburn. Partec flowcytometry system and triple color flowcytometry reagents [Dako Co], were used to evaluate peripheral blood lymphocytes population of patients admitted at the Motahary Burn Center in Tehran. Compare to healthy controls, patients with burns have shown a significant reduction in relative number of CD3[+], CD4[+] and CD8[+] T cells at three and seven adys postburn.CD4VCD8[+] ratio were below normal range in three days and remained in normal range in seven days following injury. CD19[+] B cell populations were elevated in burn patients at both three and seven days. The number of CD 16[+]NK cells were significantly declined in three days and moderately increased on day seven, following injury. Thus, the data showed that thermal burn injury suppressed T-lymphocyte subsets proliferation in various days .In addition, all compartments of showed phenotypic changes in the 3[rd] and seventh days after burn, in different groups of age. Thermal burn injury suppressed T cell subsets proliferation on day 3 and 7 postburn, when compared to normal controls. [P

Co-administration of CpG oilgonucleotides and Chenopodium album extract reverse IgG2a/IgH1 ratios and increase IFN-gamma and IL-10 prpductions in a murine model of asthma

Asthma is a disorder of increasing severity and prevalence. Recent knowledge about the pathogenesis of asthma emphasizes its inflammatory nature. CpG oligonucleotides are a class of compounds containing motifs based on the cytosine-guanine dinucleotides [CpG-ODNs]. These motifs are suppressed in mammalian DNA. They induce inflammation in mammals characterized by the induction of T helper type 1 and regulatory responses. In this paper, the effect of CpG DNA co-administration with a homemade Chenopodium album [Ch.a] extract in a murine model of asthma is reported for the first time. Balb/C mice were sensitized using Ch.a. pollen allergenic extract plus CpG-ODNs intraperitoneally and were challenged with aerosolized allergen. Results measured included IL-10 and IFN-gamma cytokines as well as IgG subclasses. For this, splenocytes from mice treated with CpG/Ag or Ag alone, were cultured in the presence of antigen. The results showed that CpG ODN administered at the time of Ch.a sensitization, effectively increased cytokines and IgG2a/IgG1 ratios compared with those in mice treated with antigen or with PBS alone[P

Immunotherapy of chenopodium album induced asthma by intranasal administration of CpG oligodeoxynucleotides in BALB/c mice

There are many therapeutic methods for allergic conditions. CpG oligonucleotides play a critical role in immunity via the augmentation of Th1 and suppression of Th2 responses. In the present study we aimed to estimate the effectiveness of intranasal administration of CpG ODN plus Chenopodium album allergen in allergic asthma compared with the administration of allergen alone and to find out how CpG ODN therapy is useful in the treatment of allergen induced asthma. BALB/c Mice were intraperitoneally and intranasally sensitized with allergenic extract precipitated on aluminum hydroxide. Therapy with CpG/Ag was performed intranasally. After antigenic challenge, a number of Immunologic variables such as serum IgE and IgG, systemic and local IL-10 and IFN-Gamma were studied in splenocytes, and lung tissue culture supernatants, respectively. Our study indicated that intranasal administration of CpG/Ag had significant increases in both systemic and local levels of IL-10 and IFN-Gamma [p

Determination of asymptomatic Chiamydia trachomatis infections by omp1 gene based-PCR

The objective of this research was to determine the prevalence of genital C. trachomatis infection in asymptomatic women by using highly sensitive nested-polymerase chain reaction [PCR] in urine sample. One hundred-forty asymptomatic women were randomly selected from those who attended gynecology out patient department of Hazraate Rasool Hospital in Tehran. First catch urine specimen were collected from all the participants. DNA extraction was performed by means of High Pure PCR Template Preparation Kit [HPPTP] according to the manufacture's instructions. Extracted DNA was tested by omp1 gene based nested-PCR, using sets of primers to amplify C. trachomatis omp1 gene. Visualization of a 1027 bp fragment from omp1 gene in agarose gel electrophoresis was considered as a positive result. In total, 140 urines were tested for determination of C. trachomatis infection. C. trachomatis omp-1 was detected in 22.1% of cases [31/140]. The overall prevalence rates of C. trachomatis in the urine sample as determined by omp1 based nested-PCR were 4.3% in group I [age, <25 years], 12.1% in group II [age, 25-34 years], 5.0% in group III [age, 35-44 years] and 0.7% in group IV [>44 years]. The highest prevalence of C. trachomatis infection [12.1%] was seen in women aged 25-34 years. This finding was not statistically significant [p=0.710]. Also, there was not relation between C. trachomatis infection and some probable risk factors such as young age [<25 years], STD history and missing use of barrier contraceptive in this study. The prevalence of C. trachomatis infection in the women not seeking health care warrants more comprehensive study using high sensitive omp1 based nested- PCR to identify and treat a large number of infected women in Iran

Study of chenopodium album allergenic extract to induce allergic asthma in murine model

The incidence of allergic and asthmatic diseases has been continuously increased in both industrial and developing countries. Extracts from various known allergens are used for the diagnostic and therapeutic purposes. To investigate the effects of an extract prepared from Chenopodium album [Ch.A.] pollen to induce allergic asthma in BALB/C mice. BALB/C mice were sensitized by i.p. injection of Ch.A. extract and alum, and an intratracheal instillation of the extract. The bronchoalveolar lavage [BAL] fluids were obtained by cannulating the trachea and lavaging the lungs and examined for eosinophilia. Splenocytes were incubated with Ch.A. extract and cell supernatants were examined for IL-4 and IL-5 by ELISA. We demonstrated that Ch.A. extract treatment in mice increased serum levels of specific IgE and production of IL-4 and IL-5 from splenocytes. An airway eosinophilia was also demonstrated in mice. These results suggest that Ch.A. allergen extract is a potential agent in inducing characteristics of allergic asthma in a mouse model useful in investigational studies

Inhibition of IL-13 by antisense oligonucleotide changes immunoglobulin isotype profile in cultured B-lymphocytes

The link between IL-13 and bronchial hyper-responsiveness has brought this cytokine as a potential therapeutic target for asthma and allergic diseases. At the present study, we address the role of B cell derived IL-13 in the IgE and other immunoglobulin development. Antisense oligo for human IL-13 m-RNA was used to study IgE down regulation. Human B-lymphocytes were purified by positive selection using magnetic cell sorting and were cultured in the complete medium plus anti-CD40 monoclonal antibody and recombinant human IL-4. Immunoglobulin assay was performed by ELISA in the presence and absence of antisense oligonucleotide. We demonstrated that IL-13 antisense causes the decrease of IgE and increase of IgA significantly and no significant changes in IgM and IgG levels [p<0.01]. We also demonstrated that both IL-13 inhibition and IL-4 removal cause the complete blocking of IgE and significant decrease of IgM and IgG levels. Our IL-13 antisense oligo can block B-cell IL-13 productions and consequently inhibits IgE production followed by IgA class switching in vitro. We suggest that in contrast to the IL-4, IL-13 is apparently more potent in the IgE switching and has no significant role in IgG and IgM levels