Hari Mohan Saxena

Guru Angad Dev Veterinary and Animal Sciences University

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Sugandha Raj

Guru Angad Dev Veterinary and Animal Sciences University

Anju Mohan

Guru Angad Dev Veterinary and Animal Sciences University

Sabia Qureshi

Veterinary & Animal Husbandry

Vimlesh Gupta

Dr. Shakuntala Misra National Rehabilitation University

Shubhada K. Chothe

University of Pittsburgh

Mudit Chandra

Guru Angad Dev Veterinary and Animal Sciences University

Deepti Chachra

Guru Angad Dev Veterinary and Animal Sciences University

Neharika Saxena

Rajasthan University of Veterinary and Animal Sciences

Daya P Gangale

Balbir Singh

Guru Angad Dev Veterinary and Animal Sciences University

Cooperative Institutions

Guru Angad Dev Veterinary and Animal Sciences University

Indian Veterinary Research Institute

Banaras Hindu University

Jamia Millia Islamia

Veterinary & Animal Husbandry

Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir

Punjab Agricultural University

Kerala Veterinary and Animal Sciences University

Government Medical College

Indian Council of Agricultural Research

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Immunotherapy of COVID-19 with Bacille Calmette – Guerin: Where is the missing red herring?

Southern African Journal of Infectious Diseases (2021)

Hari Mohan Saxena

Coronavirus Disease 2019 (COVID-19) morbidity and mortality was found to be less severe in countries where Bacille Calmette – Guerin (BCG) vaccination of the population is carried out. Conjugating Purified Protein Derivative (PPD) onto tumour cells and injecting into BCG primed mice was found to enhance anti-tumour immune response. We had proposed earlier that in vitro activated autologous anti-tumour T-cells bearing Major Histocompatibility Complex (MHC) II on their surface, if pulsed with PPD and re-infused in a BCG – primed patient, can activate PPD – specific helper T-cells and the focused secretion of lymphokines like the IL-2 can selectively amplify the antitumor T-cell response by their proliferation and activation in a specific manner bypassing the suppression exerted by the anti-idiotypic and suppressor cells. The prime – boost strategy with the BCG–PPD system can also be applied to the immunoprophylaxis and immunotherapy of COVID-19. The autologous anti-Corona virus B and T lymphocytes can be activated in vitro by inactivated virus or mitogens like Concanavalin A to express MHC class II molecules on their surface and pulsed with PPD for carrier targeting in vivo . Such PPD – pulsed activated (MHC-II+ve) anti-viral lymphocytes if transfused back into a patient already vaccinated with BCG during childhood or primed with BCG during adulthood 2 weeks before transfusion, could lead to a high magnitude of selective in vivo amplification of specific anti-viral lymphocytes, which can mount adequate and appropriate immune response to get rid of the virus and cure the patient from COVID-19. Conjugating antigens to PPD and injecting into BCG primed humans may also be helpful for immunoprophylaxis against COVID-19. Thus, PPD may prove to be the red herring in the BCG therapy of COVID-19.

10.4102/sajid.v36i1.215

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Non - Invasive Monitoring and Assessment of Phage Therapy and Phage Targeting in Bovine Brucellosis Employing a Brucella - Specific Biomarker

ACTA SCIENTIFIC MICROBIOLOGY (2020)

Anju Mohan Hari Mohan Saxena

Brucellosis caused by Brucella organisms is a major zoonosis globally.It causes heavy losses through abortions, delayed conception and infertility in animals.Antibiotic therapy is ineffective.Once infected, the animal remains carrier and sheds bacteria in milk, semen and uterine discharges for long period.We have successfully used a lytic brucella phage for therapy of Brucellosis in adult cattle.We also targeted the phage employing the live attenuated Brucella abortus strain 19 organisms to kill the virulent Brucella residing intracellularly in phagocytes in the body.The effect of both the therapies was monitored non-invasively employing Brucella RNA in blood plasma as a Brucella -specific biomarker.A single dose of the phage alone or the therapeutic vaccine (phage pulsed S-19) could substantially reduce and finally eliminate live Brucella in the body within 3 months as evident from diminished and ultimately non -detectable RNA characteristic of Brucella abortus (223 bp amplicon) in plasma by RT-PCR.Thus, phage has a potential to cure Brucellosis and abolish carrier state in cattle and RNA can serve as a specific biomarker of live Brucella for monitoring and assessment of the efficacy of the therapy.Brucellosis can be an alternative for control of the disease.Bacteriophages are viruses that infect and multiply inside the bacteria.Lytic phages quickly reproduce within and lyse the bac-

Phage therapy

10.31080/asmi.2020.03.0678

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A new biomarker for diagnosis and monitoring of effectiveness of therapy of Brucellosis

International Journal of Infectious Diseases (2018)

Hari Mohan Saxena

Background: Brucellosis is an important zoonotic disease causing huge economic losses to livestock industry and human sufferings worldwide. Current treatment of bovine Brucellosis is ineffective and infected animals become carriers. Brucella abortus Strains S19, producing antibody alone and RB51, inducing cell mediated immunity only, are used as live attenuated vaccines. However, antibody alone is ineffective for the intracellular pathogen and RB51 is Rifampicin resistant. Both strains can infect humans and cause abortions in animals. Phage lysed bacteria are harmless and retain maximum immunogenicity than conventional killing. Methods & Materials: We did immunotherapy of bovine Brucellosis by phage lysates of RB51 (RL) and S19 (SL). A single subcutaneous injection of 2 ml of lysates SL alone or RL alone or cocktail lysate (both RL and SL) was given to brucellosis affected cattle. Antibody titers, interferon gamma production and plasma RNA levels were monitored in the treated and untreated animals at various intervals upto 90 days. Results: The SL induced antibody and RL stimulated CMI. In vitro restimulation of leukocytes from RL immunized cattle induced interferon gamma. A single subcutaneous dose of 2 ml of cocktail lysate (both RL and SL), eliminated live Brucella from Brucellosis affected cattle in 3 months with Brucella specific 223 bp amplicon undetectable in plasma by RT-PCR and blood negative for live Brucella by culture by 90 day postimmunization. Conclusion: We have successfully used plasma RNA specific for live Brucella abortus as a biomarker of Brucella for accurate diagnosis and monitoring of the efficacy of immunotherapy of Brucellosis by phage lysed Brucella abortus S19 and RB51 (lysates).

10.1016/j.ijid.2018.04.4298

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Effects of Bacteriophage Therapy on Host Immune Responses in Brucellosis Affected Cattle

ACTA SCIENTIFIC MICROBIOLOGY (2020)

Anju Mohan Hari Mohan Saxena

10.31080/asmi.2020.03.0622

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Generation of phage lysates of Brucella using Brucellaphage v1

Hari Mohan Saxena Sugandha Raj

A protocol for preparing phage lysate of Brucella organisms employing a lytic brucellaphage is given here.

Lytic cycle

10.17504/protocols.io.matc2en

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A Simple Innovative Modification to Enhance the Efficacy of the Complement Fixation Test

Advances in Animal and Veterinary Sciences (2014)

Shubhada K. Chothe Hari Mohan Saxena Mudit Chandra

The Complement Fixation Test (CFT) is a serodiagnostic test of high sensitivity commonly used in the diagnosis of infectious diseases like Brucellosis.However, the conventional CFT (cCFT) may sometimes give false negative or false positive result and may lead to incorrect diagnosis.If the results are not read within 30 minutes of finishing the test, it tends to show the signs of negativity making it difficult to find the true results.In the present study, serum samples from Brucellosis affected or healthy cattle were subjected to a modified CFT (mCFT) where, in addition to the regular procedure of the CFT, anti-hemolysin antiglobulin {i.e., [F(ab') 2 ] of antibody against rabbit IgG}, was added to facilitate the reading of the test.The innovative modification showed a clear hemolysis in negative samples as the antiglobulin binds to the haemolysin leading to pairing of the antibody molecules, enhancing the complement mediated haemolysis.The novel method could help in differentiating the positive and negative samples more explicitly and the test could be read more accurately.In the modified CFT, the results could be recorded easily even after 2 hours of finishing the test, minimizing the chances of wrong diagnosis.The modified CFT with a better readout would enable more accurate serodiagnosis of infectious diseases.

Complement fixation test

Complement

10.14737/journal.aavs/2014/2.1s.13.16

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Innovative modifications to Rose Bengal plate test enhance its specificity, sensitivity and predictive value in the diagnosis of brucellosis

Journal of Microbiological Methods (2013)

Shubhada K. Chothe Hari Mohan Saxena

Rose bengal

Complement fixation test

Direct agglutination test

Bovine brucellosis

Latex fixation test

10.1016/j.mimet.2013.12.005

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Inhibition of binding of Infectious Bursal Disease Virus to chicken bursal B-cells by treatment of B-cells with antibody against a marker of immature B-cells

Journal of Immunology and Immunopathology (2007)

Daya P Gangale Hari Mohan Saxena

Infectious Bursal Disease (IBD) is an important disease of young chickens having significant economic implications for poultry industry worldwide. The exact target molecule for attachment of IBD virus (IBDV) to chicken B-cells is not known. Three monoclonal antibodies (T7, Q11 and Q13) against markers of immature B-cells of chicken were used to treat the bursal B-cells of young chickens (6 weeks old) before incubating them with IBDV. Immunoperoxidase staining of IBDV was used to assess the level of IBDV binding to B-cells without or after treatment with any of the three antibodies. The percentages of IBDV positive B-cells without and after treatment of B-cells with the T7, Q11 and Q13 antibodies were 93.42±0.47, 89.42±0.69, 91.75±0.35 and 91.58±0.57, respectively. The differences of mean values between the groups without antibody treatment and after antibody treatment were very significant in case of Q11 and Q13 antibodies (p = 0.001) and extremely significant (p<0.0001) in case of T7 antibody. ANOVA revealed an extremely significant (F = 9.624, p<0.0001) difference among the control and the three antibodies. A comparison of means between the various groups by Tukey's HSD test revealed significant differences between the control group and the three antibodies. The results suggest that the antibody T7 may be recognizing some epitope which may possibly be the target or located on or adjacent to the site of IBDV binding on B-cells.

Infectious bursal disease

Immunoperoxidase

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Analysis of Sera from Cattle Suspected for Brucellosis by Agglutination Tests and ELISA

International Journal of Current Microbiology and Applied Sciences (2018)

Anju Mohan Hari Mohan Saxena

Brucellosis is a major bacterial zoonosis of global importance. Bovine brucellosis is endemic in all states of India. However, the exact extent of the disease occurrence in the unorganized herds is not known. The present study was undertaken for serological diagnosis of brucellosis in cattle from a local gaushala (cow shelter) employing common agglutination tests RBPT, STAT and MAT and ELISA. Out of 56 samples from cattle from the Gaushala, 37.5% samples by RBPT, 64.28% each by STAT and MAT and 49.23% samples by ELISA, respectively were found to be positive for anti-Brucella antibodies. Among the 29 suspected serum samples from the field cattle, samples found to be positive for anti-Brucella antibodies were 100% by RBPT, 79.31% by MAT and 89.65% by ELISA, respectively. Titers of anti-Brucella antibodies in the sera of cattle from Gaushala were found to be 1.747+0.45, 1.935+0.62 and 0.943+0.46 by STAT, MAT and ELISA, respectively, while those in cattle from the field were found to be 1.577+0.60 and 0.783+0.59 by MAT and ELISA, respectively.

Direct agglutination test

Zoonosis

Agglutination (biology)

10.20546/ijcmas.2018.711.044

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New marker vaccine and DIVA assays for hemorrhagic septicemia in cattle

Journal of Vaccines & Vaccination (2015)

Hari Mohan Saxena Sabia Qureshi

Diva

Immunopharmacology

Vaccine adjuvant

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