Genetic association of -1562C>T polymorphism in the MMP9 gene with primary glaucoma in a north Indian population.

MMP (Matrix metalloproteinase) 9 is reported to affect glaucoma pathogenesis by altering intraocular pressure (IOP) through its role in remodeling the extracellular matrix (ECM) in the trabecular meshwork. A genetic variant at the promoter region in the MMP9 gene (-1562C>T) has a putative role in regulating its transcription rate and hence can affect genetic predisposition to primary glaucoma. The present study examined the association of -1562C>T promoter polymorphism in the MMP9 gene with Primary Open Angle Glaucoma (POAG) and Primary Angle Closure Glaucoma (PACG) in a north Indian population. A total of 729 subjects (POAG = 224, PACG = 138 and 367 controls) were recruited for the study. Genotyping for the promoter sequence variant was done with PCR-RFLP method. Genotypic and allelic frequency distribution of the POAG and PACG data sets were compared to that of controls by chi-square test and genetic association was tested under different genetic models as implemented under PLINK. Statistically significant difference was observed in the genotype frequencies between PACG cases and controls (p = 0.030). However, in the POAG cases, this difference was only borderline (p = 0.052). Genetic model analysis, under the dominant model revealed 1.6 and 1.4 fold increased susceptibility to PACG and POAG (p = 0.012, p = 0.032) respectively. A higher frequency of CT genotype was observed in PACG as well as POAG males as compared to female subjects. According to the dominant model, CT+TT genotype conferred 1.8 fold higher risk of developing PACG among male patients as compared to the control group (p = 0.048, OR = 1.87;1.00-3.50). Current findings suggest significant association of MMP9 -1562C>T polymorphism with primary glaucoma in the targeted north Indian population and warrant further replication of the findings in other populations

Prediction of an immunogenic peptide ensemble and multi-subunit vaccine for Visceral leishmaniasis using bioinformatics approaches

Visceral Leishmaniasis (VL) is a neglected tropical disease of public health importance in the Indian subcontinent. Despite consistent elimination initiatives, the disease has not yet been eliminated and there is an increased risk of resurgence from active VL reservoirs including asymptomatic, post kala azar dermatitis leishmaniasis (PKDL) and HIV-VL co-infected individuals. To achieve complete elimination and sustain it in the long term, a prophylactic vaccine, which can elicit long lasting immunity, is desirable. In this study, we employed immunoinformatic tools to design a multi-subunit epitope vaccine for the Indian population by targeting antigenic secretory proteins screened from the Leishmania donovani proteome. Out of 8014 proteins, 277 secretory proteins were screened for their cellular location and proteomic evidence. Through NCBI BlastP, unique fragments of the proteins were cropped, and their antigenicity was evaluated. B-cell, HTL and CTL epitopes as well as IFN-ɣ, IL-17, and IL-10 inducers were predicted, manually mapped to the fragments and common regions were tabulated forming a peptide ensemble. The ensemble was evaluated for Class I MHC immunogenicity and toxicity. Further, immunogenic peptides were randomly selected and used to design vaccine constructs. Eight vaccine constructs were generated by linking random peptides with GS linkers. Synthetic TLR-4 agonist, RS09 was used as an adjuvant and linked with the constructs using EAAK linkers. The predicted population coverage of the constructs was ∼99.8 % in the Indian as well as South Asian populations. The most antigenic, nontoxic, non-allergic construct was chosen for the prediction of secondary and tertiary structures. The 3D structures were refined and analyzed using Ramachandran plot and Z-scores. The construct was docked with TLR-4 receptor. Molecular dynamic simulation was performed to check for the stability of the docked complex. Comparative in silico immune simulation studies showed that the predicted construct elicited humoral and cell-mediated immunity in human host comparable to that elicited by Leish-F3, which is a promising vaccine candidate for human VL

The genotype distribution of -1562C>T MMP9 gene polymorphisms in patients with open-angle glaucoma (POAG) and angle closure glaucoma (PACG) according to CDR and IOP.

<p>The genotype distribution of -1562C>T MMP9 gene polymorphisms in patients with open-angle glaucoma (POAG) and angle closure glaucoma (PACG) according to CDR and IOP.</p

Gender-wise comparison of POAG cases and controls.

<p>Gender-wise comparison of POAG cases and controls.</p

Distribution of allele frequency, genotype frequency and genetic model of rs3918242 among PACG cases and controls.

<p>Distribution of allele frequency, genotype frequency and genetic model of rs3918242 among PACG cases and controls.</p

Sanger sequencing chromatograms for validation of genotypes.

<p>A) Homozygous CC genotype B) Heterozygous CT genotype C) Homozygous TT genotype.</p

Digested PCR product of <i>MMP9</i> -1562C>T.

<p>Well1 = 100bp Ladder; S2,10,11,12 = Heterozygous (436/242/194bp); S1,3,4,5,6,7,8,9 = Homozygous wild (436bp); UD = Undigested PCR product.</p

Gender wise comparison of PACG cases and controls.

<p>Gender wise comparison of PACG cases and controls.</p

Distribution of allele frequency, genotype frequency and genetic model of rs3918242 among POAG cases and controls.

<p>Distribution of allele frequency, genotype frequency and genetic model of rs3918242 among POAG cases and controls.</p

Frequency distribution of males and females among POAG and PACG cases with respect to control subjects.

<p>Frequency distribution of males and females among POAG and PACG cases with respect to control subjects.</p