Pushpa Gupta

Publications (13)40.41 Total impact

• Article: Expression of CXCL10 (IP-10) and CXCL11 (I-TAC) chemokines during Mycobacterium tuberculosis infection and immunoprophylaxis with Mycobacterium indicus pranii (Mw) in Guinea pig.

  Krishna Dutta Rawat, Mamta Chahar, P V J Reddy, Pushpa Gupta, Nalini Shrivastava, U D Gupta, M Natrajan, V M Katoch, Kiran Katoch, D S Chauhan
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  ABSTRACT: Mycobacterium indicus pranii (earlier known as Mycobacterium w- Mw) has been used as an immunmodulatory agent in leprosy and tuberculosis by mediating the release of various cytokines and chemokines. CXCL10 (IP-10) and CXCL11 (I-TAC) chemokines are involved in T-cell migration and stimulation of natural killer cells in Mycobacterium tuberculosis infection. In this study, the effect of heat killed M. indicus pranii (alone and in conjunction with chemotherapy) on disease progression was determined by colony forming units (CFUs) in guinea pig lung following their aerosol infection and the expression levels of CXCL10 and CXCL11 were studied by quantitative Reverse Transcriptase Polymerase Chain Reaction (qRT-PCR) and in-situ RT-PCR. Four groups of animals included; infection only (Rv), immunoprophylaxis (RvMw), chemotherapy (RvCh) and combination of immunoprophylaxis with chemotherapy (RvChMw). In the group where immunoprophylaxis was given in combination with chemotherapy, the CFU counts reduced significantly at 4(th) week post-infection as compared to animals that received immunoprophylaxis or chemotherapy alone. At the same time, all groups of animals had elevated expression of CXCL 10 which was significantly high only in animals that received Mw with or without chemotherapy. Unlike to CXCL 10, study demonstrated suppressed expression CXCL 11 in both immunoprophylaxis as well as chemotherapy groups that became up-regulated in synergistic response of immunoprophylaxis and chemotherapy. Taken together, data indicates that the expression of CXCL10 and CXCL11 positively correlates with anti-tubercular treatment (at least with combination of immunoprophylaxis and chemotherapy). Therefore, prior immunization with Mw appears to be a good immunomodulator for release of chemokines and augments the effect of chemotherapy.
  Infection, genetics and evolution: journal of molecular epidemiology and evolutionary genetics in infectious diseases 10/2012; · 3.22 Impact Factor
• Article: Epitope based recombinant BCG vaccine elicits specific Th1 polarized immune responses in BALB/c mice.

  Aparna Josephine Christy, Karthika Dharman, Gayatri Dhandapaani, Kannan Palaniyandi, Umesh D Gupta, Pushpa Gupta, S Ignacimuthu, Sujatha Narayanan
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  ABSTRACT: Developing an efficacious vaccine is one of the highest priorities in tuberculosis research. A vaccine based on T cell epitopes representing multiple antigens is an ideal approach to generate effective cellular immunity against the disease. In the present study, we have selected four T cell epitopes from four well defined Mycobacterium tuberculosis antigens, Ag85C (Rv2903c), 10-kDa culture filtrate protein (CFP-10) (Rv3874), PPE68 (Rv3873) and INV (Rv1478). The epitope encoding genes were grafted into a Cpn 10 based epitope delivery system. The cpn 10-epitope chimeras were further cloned and expressed in BCG to obtain four rBCGs (BCG::CFP, BCG::FBP, BCG::PPE and BCG::INV). Both cellular and humoral immune responses induced by these r-BCG strains were evaluated in BALB/c mice after subcutaneous injection of a single dose of 1×10(6)CFU of the individual rBCGs. Compared to the parent BCG immunized animals the splenocytes derived from rBCG vaccinated groups showed greater antigen specific proliferation, characterized with higher IFN-γ response and reduced IL-4 secretion. Also rBCG vaccination was able to induce specific humoral immune response with an enhanced IgG2a/IgG1 ratio. The rBCGs therefore favor an epitope specific Th1 type response, which is known to be important for mycobacterial immunity. Further when two of the rBCGs (BCG::CFP and BCG::FBP) were tested for their protective efficacy both the rBCGs were comparable to BCG in a H37Rv challenge study performed in guinea pigs.
  Vaccine 12/2011; 30(7):1364-70. · 3.77 Impact Factor
• Article: Mycobacterium indicus pranii as stand-alone or adjunct immunotherapeutic in treatment of experimental animal tuberculosis.

  Jaya Faujdar, Pushpa Gupta, Mohan Natrajan, Ram Das, D S Chauhan, V M Katoch, U D Gupta
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  ABSTRACT: Mycobacterium w (M.w) is a saprophytic cultivable mycobacterium and shares several antigens with M. tuberculosis. It has shown good immunomodulation in leprosy patients. Hence in the present study, the efficacy of M.w immunotherapy, alone or in combination with multi drug chemotherapeutic regimens was investigated against drug sensitive M. tuberculosis H37Rv and three clinical isolates with variable degree of drug resistance in mice. BALB/c mice were infected with M. tuberculosis H37Rv (susceptible to all first and second line drugs) and three clinical isolates taken from the epository of the Institute. The dose of 200 bacilli was used for infection via respiratory route in an aerosol chamber. Chemotherapy (5 days/wk) was given one month after infection and the vaccinated group was given a dose of 1x107 bacilli by subcutaneous route. Bacterial load was measured at 4 and 6 wk after initiation of chemotherapy. M.w when given along with chemotherapy (4 and 6 wk) led to a greater reduction in the bacterial load in lungs and other organs of TB infected animals compared to. However, the reduction was significantly (P<0.05) more in terms of colony forming units (cfu) in both organs (lungs and spleen). M.w (as immunomodulator) has beneficial therapeutic effect as an adjunct to chemotherapy.
  The Indian journal of medical research 11/2011; 134(5):696-703. · 1.84 Impact Factor
• Article: Promiscuous peptide of 16 kDa antigen linked to Pam2Cys protects against Mycobacterium tuberculosis by evoking enduring memory T-cell response.

  Uthaman Gowthaman, Vijender Singh, Weiguang Zeng, Shweta Jain, Kaneez F Siddiqui, Sathi Babu Chodisetti, Rama Krishna Gurram, Pankaj Parihar, Pushpa Gupta, Umesh D Gupta, David C Jackson, Javed N Agrewala
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  ABSTRACT: One of the main reasons considered for BCG failure in tuberculosis-endemic areas is impediment by environmental mycobacteria in its processing and generation of memory T-cell response. To overcome this problem, we developed a unique lipopeptide (L91) by linking the promiscuous peptide (sequence 91-110) of 16 kDa antigen of Mycobacterium tuberculosis to Pam2Cys. L91 does not require extensive antigen processing and generates enduring Th1 memory response. This is evidenced by the fact that L91 significantly improved the activation, proliferation, and generation of protective T cells. Furthermore, L91 surmounts the barrier of major histocompatibility complex polymorphism and induces better protection than BCG. This peptide has self-adjuvanting properties and activates dendritic cells. Importantly, L91 activates T cells isolated from purified protein derivative-positive healthy volunteers that responded weakly to free peptide (F91). In essence, L91 can be a potent future vaccine candidate against tuberculosis.
  The Journal of Infectious Diseases 09/2011; 204(9):1328-38. · 6.41 Impact Factor
• Source

  Available from: Mairaj Ahmed Ansari

  Article: RD antigen based nanovaccine imparts long term protection by inducing memory response against experimental murine tuberculosis.

  Mairaj Ahmed Ansari, Swaleha Zubair, Anjum Mahmood, Pushpa Gupta, Aijaz A Khan, Umesh D Gupta, Ashish Arora, Mohammad Owais
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  ABSTRACT: The absence of certain genomic loci that are present in most of the virulent strains of Mycobacterium tuberculosis as well as lack of lasting memory responses are some of the major causes attributed to the non effectiveness of Bacille Calmette-Gue'rin (BCG) vaccine. Immunization schedules addressing these issues can offer better strategy for protection against tuberculosis. The immunological responses evoked upon administration of archaeosome based antigen delivery system comprising T cell antigen, Rv3619c (an ESAT-6 family protein), has been assessed against experimental murine tuberculosis in BALB/c mice. Archaeosome based subunit vaccine has been found to elicit type-1 cytokines in the immunized mice. Besides effective T cell memory response, the Rv3619c based vaccine was able to reduce mycobacterial burden in the animals challenged with Mycobacterium tuberculosis infection. The data of the present study suggest that archaeosome encapsulated RD gene products offer substantial protection against M. tuberculosis infection.
  PLoS ONE 01/2011; 6(8):e22889. · 4.09 Impact Factor
• Source

  Available from: Shweta Jain

  Article: Co-administration of IL-1+IL-6+TNF-α with Mycobacterium tuberculosis infected macrophages vaccine induces better protective T cell memory than BCG.

  Vijender Singh, Shweta Jain, Uthaman Gowthaman, Pankaj Parihar, Pushpa Gupta, Umesh D Gupta, Javed N Agrewala
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  ABSTRACT: BCG has been administered globally for more than 75 years, yet tuberculosis (TB) continues to kill more than 2 million people annually. Further, BCG protects childhood TB but is quite inefficient in adults. This indicates that BCG fails to induce long-term protection. Hence there is a need to explore alternative vaccination strategies that can stimulate enduring T cell memory response. Dendritic cell based vaccination has attained extensive popularity following their success in various malignancies. In our previous study, we have established a novel and unique vaccination strategy against Mycobacterium tuberculosis (M. tb) and Salmonella typhimurium by utilizing infected macrophages (IM). In short-term experiments (30 days), substantial degree of protection was observed. However, remarkable difference was not observed in long-term studies (240 days) due to failure of the vaccine to generate long-lasting memory T cells. Hence, in the present study we employed T cell memory augmenting cytokines IL-1+IL-6+TNF-α and IL-7+IL-15 for the induction of the enhancement of long-term protection by the vaccine. We co-administered the M. tb infected macrophages vaccine with IL-1+IL-6+TNF-α (IM-1.6.α) and IL-7+IL-15 (IM-7.15). The mice were then rested for a reasonably large period (240 days) to study the bona fide T cell memory response before exposing them to aerosolized M. tb. IM-1.6.α but not IM-7.15 significantly improved memory T cell response against M. tb, as evidenced by recall responses of memory T cells, expansion of both central as well as effector memory CD4 and CD8 T cell pools, elicitation of mainly Th1 memory response, reduction in the mycobacterial load and alleviated lung pathology. Importantly, the protection induced by IM-1.6.α was significantly better than BCG. Thus, this study demonstrates that not only antigen-pulsed DCs can be successfully employed as vaccines against cancer and infectious diseases but also macrophages infected with M. tb can be utilized with great efficacy especially in protection against TB.
  PLoS ONE 01/2011; 6(1):e16097. · 4.09 Impact Factor
• Article: Coadministration of interleukins 7 and 15 with bacille Calmette-Guérin mounts enduring T cell memory response against Mycobacterium tuberculosis.

  Vijender Singh, Uthaman Gowthaman, Shweta Jain, Pankaj Parihar, Sunil Banskar, Pushpa Gupta, Umesh D Gupta, Javed N Agrewala
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  ABSTRACT: The bacille Calmette-Guérin (BCG) vaccine renders protection against tuberculosis in childhood but not in adulthood. This may be due to its failure to induce long-lasting memory T cells. T cell memory is dependent on crucial cytokine signals during the priming phases. Therefore, coadministering the BCG vaccine with cytokines may improve its efficacy. A combination of the cytokines interleukin 7 (IL-7) and interleukin 15 (IL-15) or a combination of the cytokines interleukin 1 (IL-1), interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-alpha), which are known to influence memory T cell generation, were administered along with BCG to mice. The animals were rested for a period of 240 d before they were challenged with Mycobacterium tuberculosis. Five weeks later, they were killed to study the T cell memory response. Administration of IL-7 and IL-15, but not IL-1, IL-6, and TNF-alpha, with BCG resulted in an improved CD4 and CD8 T cell memory response. Mice injected with BCG supplemented with IL-7 and IL-15 showed enhanced T cell proliferation, T helper 1-type cytokine production, and an increased pool of multifunctional M. tuberculosis-specific memory T cells. Furthermore, there was a statistically significant reduction in the mycobacterial burden in the lungs. Our results indicate that supplementation of the BCG vaccine with IL-7 and IL-15 would substantially improve its efficacy by enhancing the T cell memory response.
  The Journal of Infectious Diseases 08/2010; 202(3):480-9. · 6.41 Impact Factor
• Source

  Available from: icmr.nic.in

  Article: In vitro effect of fluoroquinolones against Mycobacterium tuberculosis isolates from Agra & Kanpur region of north India.

  Mradula Singh, D S Chauhan, Pushpa Gupta, R Das, R K Srivastava, Prashant Upadhyay, Pushpendra Singh, K Srivastava, Jaya Faujdar, G P S Jaudaun, V S Yadav, V D Sharma, K Venkatesan, S Sachan, P Sachan, K Katoch, V M Katoch
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  ABSTRACT: Fluoroquinolones (FQs) are important drugs used for treatment of drug resistant tuberculosis and are also now being considered as first line drugs to shorten the duration of treatment of tuberculosis (TB). In order to find out useful FQs for treatment of tuberculosis, the comparative efficacy of five FQs, namely, ofloxacin (OFL), ciprofloxacin (CIP), sparfloxacin (SPX), gatifloxacin (GAT) and levofloxacin (LEVX) was studied against Mycobacterium tuberculosis (MTB) isolates obtained from both treated and untreated patients from Agra and Kanpur regions of north India. A total of 162 MTB isolates [including 110 MTB isolates obtained from untreated patients (Cat-I) and 52 isolates from treated patients (Cat-II)] were tested for their susceptibilities to FQs using standard minimum inhibitory concentration (MIC) method on Löwenstein-Jensen medium. Keeping in view the therapeutically achievable drug levels, it was found that in Cat-I 97.2 per cent (107/110) isolates were sensitive to GAT, 89 per cent (98/110) to LEVX at 1 microg/ml whereas 92.7 per cent (102/110) isolates were inhibited by OFL at 2 microg/ml and 73.6 per cent (81/110) to SPX at 0.5 microg/ml. Only 63.6 per cent (70/110) isolates were found to be sensitive to CIP at 2 microg/ml which increased to 89 per cent (98/110) at 4 microg/ml (higher than achievable peak serum level). On the other hand, among 52 isolates for Cat-II, 37 (71.2%) were found to be sensitive to GAT and 33 (63.5%) to LEVX at 1 microg/ml concentration, 28 (53.8%) to SPX at 0.5 microg/ml whereas 33 (63.5%) and 24 (46.2%) isolates were found to be sensitive to OFL and CIP at 2 microg/ml, respectively. It appears that GAT has higher activity against MTB isolates followed by OFL, LEVX and SPX whereas CIP showed the lowest activity. GAT was also found to be the most effective FQ against multi-drug resistant (MDR) isolates both from Cat-I and Cat-II patients. Thus, except CIP, other FQs showed potential to be included in the treatment regimens of tuberculosis including MDR-TB.
  The Indian journal of medical research 06/2009; 129(5):542-7. · 1.84 Impact Factor
• Article: Association of mutations in rpsL gene with high degree of streptomycin resistance in clinical isolates of Mycobacterium tuberculosis in India.

  Ram Das, Pushpa Gupta, Pushpendra Singh, D S Chauhan, Kiran Katoch, V M Katoch
  The Indian journal of medical research 02/2009; 129(1):108-10. · 1.84 Impact Factor
• Article: Determination of ethambutol MICs for Mycobacterium tuberculosis and Mycobacterium avium isolates by resazurin microtitre assay.

  G P S Jadaun, Chhaya Agarwal, Hirdesh Sharma, Zafar Ahmed, Prashant Upadhyay, Jaya Faujdar, Anuj Kumar Gupta, Ram Das, Pushpa Gupta, D S Chauhan, V D Sharma, V M Katoch
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  ABSTRACT: To test susceptibilities of Mycobacterium tuberculosis (MTB) isolates to ethambutol by the Löwenstein-Jensen (LJ) proportion method and resazurin microtitre assay (REMA) and to evaluate REMA for the determination of ethambutol MICs for MTB and Mycobacterium avium isolates. A total of 50 MTB and 20 M. avium isolates were tested to determine the MICs of ethambutol by REMA and agar dilution method. MTB isolates were also tested by the LJ proportion method. REMA provided ethambutol susceptibility results for all the isolates within 8-9 days. For MTB isolates, REMA showed 96.7% sensitivity, 100.0% specificity and 98.0% accuracy when LJ proportion results were taken as 'gold standard'. For both MTB and M. avium isolates, the MICs determined by REMA were lower than those determined in agar medium, indicating that MIC values determined by REMA are closer to the actual MICs for the isolates. REMA can be used as a rapid and inexpensive method for mycobacterial drug susceptibility testing against ethambutol. In comparison with the agar method, the MICs determined by REMA can more accurately be correlated with achievable plasma concentrations of antimycobacterial agents.
  Journal of Antimicrobial Chemotherapy 08/2007; 60(1):152-5. · 5.07 Impact Factor
• Source

  Available from: Ram Das

  Article: Simultaneous ethambutol & isoniazid resistance in clinical isolates of Mycobacterium tuberculosis.

  Pushpa Gupta, G P S Jadaun, Ram Das, U D Gupta, Kiran Srivastava, Aradhana Chauhan, V D Sharma, D S Chauhan, V M Katoch
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  ABSTRACT: There is a need to understand the nature of drug resistance patterns and predictors of emergence of drug resistance in Mycobacterium tuberculosis. There could be common factors/mechanisms for resistance to the drugs, isoniazid and ethambutol, both acting on cell wall. The present study was conducted to analyze the antimycobacterial susceptibility patterns of M. tuberculosis isolates to determine the minimum inhibitory concentrations (MICs) of ethambutol for M. tuberculosis; and to find out possible association of ethambutol resistance with isoniazid resistance. A total of 380 M. tuberculosis isolates were tested for their susceptibilities to ethambutol at 2, 4, 6 microg/ml, isoniazid at 1 microg/ml and rifampicin at 64 microg/ml using MIC method. 44.21, 24.73 and 14.21 per cent isolates were resistant to ethambutol at concentrations of 2, 4 and 6 microg/ml respectively. At 6 microg/ml of ethambutol concentration, 85.18 per cent ethambutol resistant isolates were resistant to isoniazid also. At the same ethambutol concentration a fraction of 28.75 per cent isoniazid resistant isolates were ethambutol resistant. Ethambutol resistance was accompanied with isoniazid resistance in a large percentage of isolates whereas ethambutol resistance was weakly linked with multidrug resistance. On the other hand, association between isoniazid and ethambutol resistance was weak showing one way linkage.
  The Indian journal of medical research 03/2006; 123(2):125-30. · 1.84 Impact Factor
• Article: Sexual risk behaviors in HIV positive individuals in north India.

  Eshagh Dortaj Rabori, Pushpa Gupta, R M Pandey, D Chattopadhaya, Shiv Lal, Usha K Baveja
  The Journal of communicable diseases 10/2005; 37(3):249-53.
• Article: Risk taking behavior and Sexually Transmitted Diseases: a study among men.

  Pushpa Gupta, Arun Kr Sharma, V G Ramachandran
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  ABSTRACT: Sexually Transmitted Diseases (STDs) among men not only jeopardize their own health but also increase sexual morbidities among their spouses. STDs are primarily attributed to high-risk sexual behavior. The study was carried out among male patients attending the OPD of a Government dispensary. Risk taking behavior was assessed using a structured, pre-tested questionnaire and STDs were diagnosed using syndromic approach supported by laboratory investigations. Three hundred two men were selected by systemic random sampling for inclusion in the study. 39% had pre-marital sexual relationship, 20% had sex with Commercial Sex Workers. 12% of the married men had extramarital sex mostly with CSWs. 27% had more than one sex partner ever. Only 3.6% used condoms consistently. Thirteen per cent respondents had history of symptoms suggestive of STDs. Prevalence of syphilis, gonorrhea and other STDs among study subjects were 3.6%, 0.7% and 0.7% respectively. High risk sexual behavior is widely prevalent and STDs are also common. Behavior change communication and early diagnosis and prompt treatment of STDs will reduce the burden significantly.
  The Journal of communicable diseases 04/2005; 37(1):51-7.