Immunogenicity and protective efficacy of 3A truncated negative marker foot-and-mouth disease virus serotype A vaccine.

Foot-and-mouth disease (FMD) is a highly contagious, economically significant disease of cloven-hoofed animals caused by FMD virus (FMDV) of the Picornaviridae family. Vaccination of susceptible animals with inactivated virus vaccine is the standard practice for disease control. The prophylactic use of the inactivated vaccines has reduced the disease burden in many countries endemic to FMD. In the process of implementation of the mass vaccination program and disease eradication, it is essential to differentiate infected from vaccinated animals (DIVA) where a large proportion of the animal population is vaccinated, and disease-free zones are being established, to help in sero-surveillance of the disease. In such a scenario, the use of a negative marker vaccine is beneficial to rule out false-positive results in a disease-free zone. Here we report the construction and rescue of an infectious cDNA clone for FMDV serotype A Indian vaccine strain lacking 58 amino acid residues (87-144 amino acid position) in the carboxy-terminal region of the viral 3A protein. The recombinant deletion mutant virus showed similarity in the antigenic relationship with the parental strain. Immunization of guinea pigs with the inactivated vaccine formulated using the deletion mutant virus induced potent immune response with 100% protective efficacy upon challenge with homologous virus. Further, we show that sera from the guinea pigs infected with the deletion mutant virus did not show reactivity in an indirect ELISA test targeting the deleted portion of 3A protein. We conclude that the recombinant deletion mutant virus vaccine along with the newly developed companion indirect ELISA targeting portion of FMDV 3A protein could be useful in the implementation of a precise DIVA policy in our country when we reach FMD free status with vaccination.

Fever with Rash, an Alarm to the Physicians - A Case Report of Atypical Measles.

Measles is a highly contagious disease caused by Paramyxovirus group. It has been estimated that globally deaths occurs out of measles. The disease still remains as one of the leading cause of death among young children and adults despite the effective vaccination. The outbreak of measles is on rise in recent years with serologically confirmed cases in India. The incidence and complications of atypical measles is usually severe among older age groups even prior immunization. We report a case of 15-year-old male with symptoms of fever and rash with possible differential diagnosis and emphasize on the clinical presentation of atypical measles among adolescent age group with an additional evidence of investigations.

Comparative efficacy of virus like particle (VLP) vaccine of foot-and-mouth-disease virus (FMDV) type O adjuvanted with poly I:C or CpG in guinea pigs.

Foot-and-mouth disease (FMD) is one of the most contagious and economically important diseases of cloven-hoofed livestock. Currently used inactivated FMD vaccines have short lived immunity besides risk of handling live virus. We studied recombinant FMD virus like particles (VLPs) encoded by FMDV type O/IND/R2/75 polyprotein genes expressed in Sf9 cells and adjuvanted with CpG or Poly I:C in inducing protective immune response in guinea pigs. Guinea pigs immunized with VLP + CpG vaccine had shown markedly higher cell mediated immunity (CMI) in comparison to the conventional vaccine group as evident from higher levels of IgG2 than IgG1. Although the humoral response was less in VLP + CpG compared to conventional vaccine, the lymphocyte stimulation index was more in VLP + CpG compared to conventional and VLP + Poly I:C vaccine groups. Finally the challenge experiments on 28 and 56 dpv had shown 75% protection in VLP + CpG immunized guinea pigs primary and boosted animals, while 50% and 62% protection in VLP + Poly I:C in primary and boosted animals, respectively. In conclusion, CpG adjuvant was found to be superior followed by ISA206 and Poly I:C in eliciting protection in VLP based FMD vaccines in guinea pigs.

Protective immune response to liposome adjuvanted high potency foot-and-mouth disease vaccine in Indian cattle.

BACKGROUND: Foot-and-mouth disease (FMD) vaccines applied for prophylactic use in endemic areas provide short-lived immunity requiring regular boosters. Indian FMD control program recommends twice a year vaccination. Development of high potency vaccines that provide better immune response can singificantly contribute to control programme by reducing the frequency of vaccination. The present study explores new adjuvants to enhance the protective efficacy of inactivated trivalent FMD vaccines. METHODOLOGY AND PRINCIPAL FINDINGS: VacciMax(®) is a novel adjuvant which uses a liposome-based oil emulsion platform. Cattle were immunized using VacciMax-A and VacciMax-B FMD vaccines and evaluated for protective efficacy. Similar groups of animals were also boosted after 6 months to study the effect of booster immunisation on protection against homologous challenge. Serum samples from immunized animals were tested by virus neutralization test (VNT) and liquid phase blocking ELISA (LPBE). After challenge, animals were screened for virus load by real-time PCR and reactivity in non-structural protein (3ABC) antibody detection ELISA to corroborate the protection data. A single dose of VacciMax-A formulation elicited higher percentage protection (63%) in VacciMax-A compared to 25% in VacciMax-B upon challenge at one-year post-vaccination. Upon boosting at 6 months also, VacciMax-A group showed higher levels of protection (100%) compared to VacciMax-B (86%), even though both the groups elicited comparable VNT titre (p=0.4964). The results also demonstrated that intramuscular route was preferrable over subcutaneous route of administration. CONCLUSION: The study demonstrates that immunization with VacciMax-A-IM adjuvanted FMD vaccine with high antigen payload under boosting regimen could effectively be used as potent vaccine to maintain herd immunity.

Rescue of infective virus from a genome-length cDNA clone of the FMDV serotype O (IND-R2/75) vaccine strain and its characterization.

We report here the construction and characterization of an infectious cDNA clone of the Indian vaccine strain of foot and mouth disease virus (FMDV) serotype O, IND-R2/75. Viral genome was amplified by reverse transcription-polymerase chain reaction (RT-PCR) in five fragments and subsequently assembled sequentially in a plasmid vector to generate a complete cDNA clone, flanked by the T7 RNA polymerase promoter and poly (A) tail at 5' and 3' ends, respectively. Transfection of BHK-21 cells with the RNA transcribed from this genome-length cDNA construct allowed the recovery of infectious recombinant FMDV particles as evidenced by cytopathic effect in BHK-21 cells. Characterization of the recombinant virus revealed its similarity to the parental strain. Recombinant virus could be distinguished from the parental virus based on the presence of a unique marker sequence in the former, which was incorporated in the cDNA using a silent mutation. The virus showed no significant amino acid changes in the capsid-coding region when serially passaged up to ten times in BHK-21 cells, while retaining the marker sequence.

Prevalence of peste des petits ruminants among sheep and goats in India.

This study measured the clinical prevalence of peste des petits ruminants (PPR) among sheep and goats in India between 2003 and 2009 by analyzing clinical samples from suspected cases of PPR that were submitted to the Rinderpest and Allied Disease Laboratory, Division of Virology, IVRI, Mukteswar for PPR diagnosis. PPR outbreaks were confirmed by detecting PPR virus (PPRV)-specific antigen in the clinical samples. Clinical samples (blood, nasal swabs, spleen, lymph node, kidney, liver, intestine, and pooled tissue materials) were taken from a total of 592 sheep and 912 goats in different states of India and screened for the presence of PPRV antigen using a monoclonal antibody based sandwich ELISA kit. A total of 20, 38, and 11 laboratory-confirmed PPR outbreaks occurred among sheep, goat, and combined sheep and goat populations, respectively. Our findings provide evidence of widespread PPR endemicity in India. The underlying reasons could be variations in husbandry practices in different geographical regions, agro-climatic conditions, and livestock migration. Furthermore, decrease in the number of PPR outbreaks over time might be due to the effectiveness of current live PPR vaccines and timely vaccination of target species. Vaccination against PPR has been practiced in India since 2002 to control this disease.

Vaccines against peste des petits ruminants virus.

Peste des petits ruminants (PPR) is a highly contagious and economically important viral disease affecting goats, sheep and wild ruminants. The disease is currently circulating in Asian and African countries, creating problems in small ruminant farming. Current control of the disease mainly includes isolation and disinfection of the contaminated environment, and administration of a live-attenuated vaccine, which provides a strong immunity. Maintenance of cold chain for vaccine efficacy has proven difficult in subtropical countries. A thermostable live-attenuated conventional or recombinant vaccine is a way to avoid cold chain-associated problems in tropical and subtropical countries. Mass vaccination of sheep and goats in endemic countries might be a pragmatic approach to control PPR in the first phase of disease eradication. In the future, the development of a marker vaccine with a robust companion test may help in serosurveillance to identify infection in vaccinated animals to control PPR disease. Here, we discuss available potent PPR vaccines and the future possibility of developing new-generation vaccines against PPR.

Possible control and eradication of peste des petits ruminants from India: technical aspects.

The peste des petits ruminants (PPR) is an acute and highly contagious, notifiable viral disease of sheep and goats that causes substantial morbidity and mortality. There are three cell culture-based live attenuated PPR vaccines available (one from an African isolate and two from Indian isolates). The PPR vaccine produced by the Indian Veterinary Research Institute has been extensively evaluated in the field and found safe and potent in sheep and goats in India. Diagnostic tests, such as the sandwich enzyme-linked immunosorbent assay (s-ELISA), competitive ELISA, single and duplex reverse transcriptase-polymerase chain reactions (RT-PCRs) and RT-PCR-ELISA at the Indian Veterinary Research Institute have also been validated on a large scale. Furthermore, the expertise that remained after the successful eradication of rinderpest in the National Project on Rinderpest Eradication can be utilised effectively for the eradication of PPR without much additional budgetary expense. Thus, the availability of an effective vaccine, accurate diagnostic tests for PPR and an experienced infrastructure prompt us to propose a national project for a peste des petits ruminants eradication programme on the lines of National Project on Rinderpest Eradication. This would greatly enhance the prospects of PPR eradication not only on a national level but also from the Asian continent, alleviate poverty and, in turn, contribute to the national economy.