HLA-DR2 subtypes & immune responses in pulmonary tuberculosis

Background & objectives: HLA-DR2 has been shown to be associated with the susceptibility to pulmonary tuberculosis and altered antibody and lymphocyte response in pulmonary tuberculosis. In the present study, the influence of DR2 subtypes on antibody titre and lymphocyte response ,to Mycobacterium tuberculosis culture filtrate antigens (10 μg/ml) was studied in 22 patients with active pulmonary TB (ATB), 50 inactive (cured) TB (ITB) patients and 36 healthy control subjects. Methods. HLA-DR2 gene was amplified by polymerase chain reaction (PCR) and dot-blotted. Genotyping of DRBl*1501, *1502, *1503, *1601 and *1602 was carried out using sequence specific oligonucleotide probes (SSOPs) and detected by chemiluminescence method. Antibody titre as well as lymphocyte response to M.tuberculosis antigens were measured by enzyme linked immunosorbent assay (ELISA) and lymphocyte transformation test (LTT) respectively. Results: The allele frequency of DRB1*15Ol was significantly increased in pulmonary tuberculosis patients as compared to controls (P<0.05). No marked difference in the antibody titre and lymphocyte response to M. tuberculosis antigens was observed between the DRBl *1501, *1502 and *1503 positive or negative controls, ATB and ITB patients. DRBl *1501 and *1502 positive as well as negative ATB patients showed a higher antibody titre as compared to controls and ITB patients. ITB patients with *1502 showed a higher lymphocyte response as compared to *1502 positive controls (P<0.001) and ATB patients (P<0.05). Similarly, an increased lymphocyte response was observed in *1501, and *I503 negative ITB patients compared to *1501 and *1503 negative controls and ATB patients. Interpretation & conclusion: The present study revealed that DRBl *1501 may be associated either alone or with other DR2 alleles, with the susceptibility to pulmonary tuberculosis. None of the DR2 alleles influenced the antibody and lymphocyte response to M tuberculosis culture filtrate antigens. This suggested that HLA-DR2 gene/gene products as a whole may influence the immune response in pulmonary tuberculosis

Vitamin D receptor and interleukin-1 receptor antagonist gene polymorphism in spinal tuberculosis

Our earlier studies revealed that both MHC (Major Histocomptibility Complex) and non-MHC genes are associated with the susceptibility to pulmonary tuberculosis (TB). To find out whether non-MHC genes such as vitamin D receptor (VDR) and interleukin-1 receptor antagonist (IL-1RA) genes are associated with the susceptibility to spinal TB (extrapulmonary form of TB), the present study was carried out in spinal TB patients (n=66) and spouses of TB patients (spinal-TB and pulmonary-TB) ( n = 80) (family contacts). A trend towards an increased per cent genotype frequency of IL-1RA genotype variant 22 (12.1%) was seen in spinal TB patients when compared to the controls (3.8%) (spouses of the patients) (P=0.057; odds ratio 3.5). No difference was observed in the frequency of VDR genotypes between the overall spinal TB patients and the family contacts. However, the VDR mutant genotype tt was seen at a higher frequency in female patients with TB spine (TBS) (12.8%) than female contacts (4.2%) ( P >0.05 not significant; odds ratio 3.5). Among the contacts, a significantly increased frequency of wild type genotype TT (wild homozygotes) was seen in female contacts (55.1%) than male contacts (16.1%) (P =0.0012). Similarly a significant decrease in tt genotype was seen in female contacts (4.1%) than male contacts (25.8%) (P=0.012). The present study suggests that IL-1RA genotype 22 may be associated with the susceptibility to spinal TB. Moreover, vitamin D receptor tt genotype may be associated with the susceptibility to spinal TB in female patients. The study reveals that multicandidate genes may be associated with the susceptibility to spinal TB

HLA-DR phenotypes and lymphocyte response to M. tuberculosis antigens and in cured spinal tuberculosis patients and their contacts

Background: Our earlier studies on Human Leucocyte Antigens (HLA) in pulmonary tuberculosis patients revealed the association of HLA-DR2 antigen with susceptibility to pulmonary TB and DR2 antigen has been shown to influence the immunity to tuberculosis. Objectives: The present study was carried out to find out whether HLA-DR antigens are associated with susceptibility to spinal tuberculosis. Moreover, the role of HLA-DR antigens on lymphoproliferative response to Mycobacterium tuberculosis culture filtrate antigens was studied using Lymphocyte Transformation Test (LTT). Material and Methods: HLA-DR genotyping and lymphoproliferative response was carried out in 63 cured spinal TB patients and 63 control subjects (spouses of pulmonary and spinal TB patients). Results: A trend towards an increased frequency of HLA-DR9 antigen was observed in spinal TB patients compared to controls. A significantly decreased lymphocyte response to M. tuberculosis antigens was observed in HLA-DR9 antigen positive control subjects compared to HLA- DR9 antigen negative subjects (P=0.0009) whereas increased response was observed with DR9 positive cured spinal TB patients compared to HLA-DR9 antigen negative patients. Further, HLADR3 antigen positive patients showed a decreased lymphocyte response compared to HLA-DR3 antigen negative patients (P<0.05). Conclusion: The study suggests that HLA-DR9 antigen either alone or in combination with other HLA antigen as lhplotype and non-HLA genes may be associated with susceptibility to spinal TB and play a regulatory role on the immune response to M. tuberculosis in spinal tuberculosis patients

Influence of non-MHC genes on lymphocyte response to Mycobacterium tuberculosis antigens and tuberculin status in Pulmonary tuberculosis

Background & objectives : Major histocompatibility complex (MHC) genes are known to influence the immune functions. In the present study, the influence of non-MHC genes such as mannose binding protein (MBP), vitamin D receptor (VDR) and interleukin-1 receptor antagonist (IL-IRA) gene polymorphisms on lymphocyte response to Mycobacterium tuberculosis culture filtrate antigen (10 μg/ml) was studied in 44 patients with active pulmonary TB and the family contacts (35) and in 32 normal healthy subjects. The influence of these gene polymorphisms on tuberculin (1TU of PPD of M. tuberculosis) reactivity status in 146 pulmonary TB patients was also studied. Methods : The MBP and VDR genes were amplified using polymerase chain reaction (PCR) and genotyping was carried out using sequence specific oligonucleotide probes by dot blot and IL-1RA by agarose gel electrophoresis. Results : A significantly decreased lymphocyte response to M. tuberculosis antigen was seen in pulmonary TB patients positive for functional mutant homozygotes of MBP (00) compared to heterozygote carriers (AO; P<0.02) and wild homozygotes (AA; P<0.01). The variant mutant genotype (tt) of VDR gene was associated with an increased lymphocyte response in control subjects compared to active TB patients with tt genotype (P<0.05). Heterozygote carriers of MBP (AO) were associated with a significantly (P<0.001) decreased tuberculin reactivity compared to wild homozygotes (AA). The VDR genotype Tt (heterozygote carrier) was associated with an increased tuberculin reactivity in female TB patients as compared to male patients (P<0.001). Interpretation & conclusions : The present study suggested that MBP and VDR genes influence the cell mediated immune response in pulmonary TB patients. Non-MHC genes along with HLA-Class II genes/gene products may be playing an immunoregulatory role in the mechanism of susceptibility/resistance to tuberculosis

Association of vitamin D receptor gene variants of BsmI, ApaI and FokI polymorphisms with susceptibility or resistance to pulmonary tuberculosis

Vitamin D receptor (VDR) gene polymorphism was studied to find out whether the variants of this gene are associated with susceptibility or resistance to pulmonary tuberculosis (PTB) and bacteriological relapse of tuberculosis. BsmI, ApaI and FokI polymorphisms of VDR gene were studied in PTB patients (n = 120), patient contacts (spouses of the patients; n = 80), bacteriological relapse patients (n = 48) and quiescent patients (n = 48). Significant increase of Bb genotype (heterozygote carrier) of BsmI polymorphism (P = 0.028) and FF genotype (homozygotes of common allele F) of FokI polymorphism (P = 0.034) were observed in male PTB patients than male contacts. The BB genotype (homozygote of common allele B) of BsmI polymorphism and AA genotype (homozygote of common allele A) of ApaI polymorphism were increased in male contacts than male PTB patients (BB: P = 0.018; AA: P = 0.04). No significant differences were found among female patients and female contacts. In bacteriological relapse cases of PTB, a decreased frequency of AA genotype (P = 0.015) and an increased frequency of Aa genotype (P = 0.024) were observed in bacteriological relapse patients than quiescent patients of PTB. The present study suggests that Bb genotype of BsmI polymorphism and FF genotype of FokI polymorphism of VDR gene may be associated with the susceptibility to tuberculosis in males. The BB and AA genotypes may be associated with resistance to PTB in males. The genotype Aa may be associated with bacteriological relapse and AA may be associated with protection against bacteriological relapse

Effect of vaginal pH on efficacy of dinoprostone gel for labour induction

Background: Induction of labour is defined as an intervention designed to artificially initiate uterine contractions leading to progressive dilatation and effacement of the cervix and birth of the baby. The aim of the study was to evaluate the influence of vaginal pH on the efficacy of dinoprostone gel for labor induction.Methods: A prospective study conducted on 200 subjects within 1 year in India. The Bishop score and vaginal pH (with pH paper, Sigma Chemical Company, St. Louis, USA) of subjects undergoing induction of labor with dinoprostone gel was assessed prior induction. After 6 hours of induction, PGE2 gel was repeated or labor was augmented. The vaginal pH measured was compared with age, parity, Bishop Score, time to enter into active phase of labor, and the mode of delivery. The significance of association was calculated by Chi-square test.Results: Majority subjects had pH in the range 5-6.Subjects with higher parity were associated with higher vaginal ph. Higher vaginal pH was associated with a higher Bishop score prior to induction, responded to single induction, and had a higher  number of vaginal deliveries than those with lower vaginal pH. There was no significant association found in vaginal pH and the time taken to enter into active phase of labor. Subjects with pregnancy induced hypertension were found to have higher vaginal ph.Conclusions: Parity influences vaginal pH and vaginal pH itself has a significant effect on the Bishop Score prior induction. Hence knowing the vaginal pH prior induction could be a useful tool to assess the labor outcome in induction with PGE2

Effect of Organic Nutrition Practices on Papaya (cv. Surya) Fruit Yield, Quality and Soil Health

A field experiment was conducted during 2005-07 at Indian Institute of Horticultural Research, Bangalore, on papaya cv. Surya with six organic treatments along with recommended dose of fertilizers and no manure/fertilizer application. Results indicated that crop growth and fruit yield were higher in inorganic fertilizer treatment (55 t ha1) compared to organic treatments (26.9 to 38 t ha-1). There was no significant variation in average fruit weight and TSS, but shelf life of the fruit was significantly higher in organic treatments (6.2 to 7.9 days) as compared to inorganic fertilizer treatment (5.1days). Among the treatments, application of 7 kg urban compost plant-1 or 10 kg FYM plant-1 was found to be ideal for improving soil health in terms of microbial population, and biochemical reaction compared to other treatments

NRAMP1 gene polymorphism in pulmonary and spinal tuberculosis

NRAMP1 gene polymorphisms such as 823 C/T (exon 8), deletion of TGTG in the 3'-UTR (3¢ untranslated region) and D543N G/A (exon 15) were studied to find out whether the gene polymorphic variants are associated with the susceptibility or resistance to tuberculosis (TB). The study was carried out in pulmonary (n = 100) and spinal TB patients (n = 57) and control subjects (n = 112). No difference was observed in the variant genotype frequencies of NRAMP1 – 823 C/T, TGTG+/del and D543N G/A polymorphisms. However, a trend towards an increased frequency of the variant genotype 823 C/T (heterozygotes) was observed in pulmonary TB patients than control and spinal TB patients (odds ratio (OR) : 1.6; confidence interval (CI): 0.74–3.4). A trend towards an increased frequency of the variant genotype TGTG del/del of 3'-UTR was observed in control subjects than pulmonary and spinal TB patients (OR : 0.48; CI: 0.10–1.78; OR: 0.42; CI: 0.04– 2.12, respectively). The above trends were not significant. The study suggests that NRAMP1 gene may not be associated with the susceptibility to pulmonary and spinal TB in the Indian population. Major Histocompatibility Complex (MHC) and other non- MHC gene polymorphic variants may be associated

Immune response to Mycobacterium tuberculosis culture filtrate antigen in cured spinal tuberculosis patients and their spouses

Humoral and cell mediated immune responses were studied in cured spinal tuberculosis patients and their spouses to understand immunit to tuberculosis in cured patients. Antibody litre and immune complex levels were measured and lymphocyte response to Mycobaclerinm tuberculosis culture filtrate antigen was observed in cured spinal tuberculosis patients ( n = 3 0 ) and their spouses (n =27). A trend towards increased antibody litre was seen in cured patients as compared to their spouses. Significantly increased circulating immune complex levels, as measured by PEG OD280 (polyethylene glycol optical density 280) were seen in the contacts compared to cured patients. And a trend towards increased lymphocyte response to Mtuberculosis culture filtrate antigen was seen with different antigen concentrations (0.1,1 and 10 μg /ml). Moreover, the effect of active-pulmonary-Tuberculosis (AT B) plasma taken from 1ILA-DR2 positive and DR2 negative patients on lymphocyte response of the cured patients showed no dramatic immunomodulatory effect in the lymphocyte response when treated with DR2 positive or DR2 negative plasma. The study suggests that the memory response lo Mtuberculosis is well maintained even after 10-15 years of treatment