5^th Asia Pacific Global Summit and Expo on Vaccines & Vaccination July 27-29, 2015 Brisbane, Australia

Jan Fohlman has completed his PhD and MD from Freiburg, Linköping, Uppsala University and has been working as a consultant in infectious diseases for >30 years, now at Dept of Research and Development Region Kronoberg, Växjö, working closely with Linnaeus Univeristy. He is a board member of 3Diva AB, a recently started vaccine development company . He has published more than 100 papers in reputed journals and has been serving as a referee for multiple journals.

Background: Attempts to conquer influenza are struggling with the complexity of hypervariability. CDC issued a warning already in December 2014 that the profile of influenza viruses currently circulating (with A/H3N2 predominating) did not match with the corresponding strain in the vaccine. The flu season 2015 was severe. There was no protection against 97 % of the virus strains, 40 % higher mortality in England, 7 % of all mortality in USA in December due to influenza and about 1000 persons succumbing to flu in Sweden this season.
Aim: This raises the question whether it is possible to construct a vaccine that gives a heterotypic protection. ISCOM H1N1 influenza vaccine protected mice against H2N2 influenza strain challeng(1). A commercial vaccine (Pandemrix®) was issued in 2010 and gave a longer lasting immunoprotection, apparently due to the use of squalene as an adjuvant. Unfortunately a strain of influenza was selected that later turned out to cause narcolepsy in predisposed individuals, esp children (3). This could not have been anticipated and not even tested beforehand, due to the rarity of the condition, even if it was increased about 3-fold.
Methods: A further development of ISCOM is now under investigation under the name of G3/DT®. A vaccine H1N1 A/California/7/2009 (H1N1pdm09) protected against a lethal challenge with antigenically distinct H1N1 A/PR8/34 in a mouse model (2). The adjuvanted vaccine gave a high level of protective antibodies. Also influenza A virus-specific CD8+ T lymphocytes were induced and seem responsible for the heterotypic response.
Conclusions: No adjuvanted influenza vaccine has so far been licensed in the US. To meet the problem of non-matching strains adjuvanted vaccines might be advantageous for protection against yearly flu infection.

Hiroko Toriniwa recieved her phD frome Faculty of Environmental Earth Science, Hokkaido University. She is Senior Researcher of Vaccine Research Laboratories, R&D Division, KITASATO DAIICHI SANKYO VACCINE CO., LTD. She have researched development of Japanese encphalitis virus vaccine. She is coauthor of the patent : Method of producing japanese encephalitis vaccine stably storable over long time and use of the vaccine with reference PCT/JP2008/073732.

We previously developed an inactivated Japanese encephalitis virus (JEV) vaccine (ccJE) using serum-free cultured Vero cells. The immunogenicity of this ccJE vaccine was enhanced by formulation with AdvaxTM, a novel polysaccharide adjuvant based on delta inulin thereby enabling JEV protection after just a single immunisation. Advax adjuvant potently stimulated humoral and cellular immunity but avoided the reactogenicity of other adjuvants. Immunisation of mice with the ccJE-Advax vaccine combination induced cross-neutralising antibodies against other Flaviviruses belonging to the JEV serocomplex, including Murray Valley encephalitis virus (MVEV) and West Nile virus (WNV). We thereby sought to identify the immune mechanism underlying this cross protection. The ccJE-Advax combination changed the balance of T cell immunity as reflected by changes in the IgG antibody isotypes. Splenocytes from mice immunized with ccJE-Advax when compared to mice immunized with ccJE antigen alone exhibited increased IFN-γand IL-17 production when stimulated with ccJE antigen, consistent with Advax adjuvant increasing Th1 and Th17 immunity. Increased antigen-specific T-cell IL-17 production and protection against JEV challenge was also seen in an IFN-γ knockout mouse model, indicating that IFN-γwas not critical to ccJE-Advax mediated protection. Thus the combination of ccJE-Advax induces broad humoral and cellular immunity against JEV, translating into single-dose protection. This broader immunity obtained through use of Advax adjuvant in the vaccine enables enhnhanced cross-neutralising antibody and T-cell responses against not just JEV but also other JEV serocomplex flaviviruses including WNV and MVEV, with enhanced antigen-specific Th1 and Th17 immune responses.

Khairunnisa graduated with a Diploma in Pharmaceutical Sciences and Technology from Temasek Polytechnic, Singapore in 2010. Upon graduating, she obtained a Bachelor of Science (Biochemistry) with Honours Class I from the University of Queensland, Australia in 2012. She is currently holding a University of Queensland International Scholarship for her postgraduate studies. During her studies, she has received various travel awards to attend workshops and conferences abroad. Currently, she is finilizing her PhD under the supervision of Prof. Istvan Toth and Dr. Mariusz Skwarczynski at the University of Queensland.

For the past decade, Group A Streptoccocus (GAS) has been one of the top ten pathogens causing mortality and morbidity. Treatment with antibiotics has proven to be successful; however,c the lack of early medical attention could lead to invasive GAS infection. In addition, rheumatic heart disease, dubbed as the ‘silent killer’, is the most critical sequelae of GAS, claiming more than 1 million lives each year.1 Our vaccine strategy utilizes the lipid core peptide (LCP) as a self-adjuvanting system2 in conjunction with liposomes as a delivery system. Liposomes are known for their efficacy in delivering antigens as well as for their targeting capability.3,4 Two epitopes were used, a) J14, KQAEDKVKASREAKKQVEKALEQLEDKVK, derived from the surface associated M protein of GAS; b) P25, KLIPNASLIENCTKAEL, a universal T-helper epitope. Seven different peptides (branched and linear), were synthesized using standard solid phase peptide synthesis and purified (≥95%) to attain moderate to high yields (44-80%). Liposomes were formulated giving an average size of 170 nm and a zeta potential of +57 mV. All compounds had varying percentages of encapsulation efficacy ≥65%. The lead vaccine candidate (liposomes encapsulating P25-K(J14)-LCP) had the highest encapsulation efficacy of 98%. Mice immunized intranasally with the lead vaccine candidate elicited the highest IgG and IgA antibody titres against J14 compared to all vaccine candidates and positive controls which included known adjuvant, cholera toxin B subunit. In addition, mice also retained high levels of serum IgG five months post final immunization. In conclusion, we prepared the first LCP-nanoliposome-based peptide vaccine candidate against GAS.

Noushin obtained her Master of Science (First Class Honors) in Shahid Chamran University, Iran in 2010. She has three years of work experience in the field of virology and vaccine design. She has also received various travel awards to attend workshops and conferences. Currently, she is holding a University of Queensland International Scholarship for her PhD study. She is now a second year PhD student under the supervision of Prof. Istvan Toth, Prof. Paul Young and Dr. Mariusz Skwarczynski at the University of Queensland.

Respiratory syncytial virus (RSV) is a leading cause of lower respiratory tract infection in infants and young children. RSV also causes serious illness in the elderly, immuno-compromised and persons with cardiopulmonary diseases.1 The significant morbidity and mortality associated with RSV infection emphasises the urgent need for vaccines against RSV infection. The failure of formalin-inactivated RSV clinical trials in the 1960s2 directed researches towards finding new approaches for the development of RSV vaccines. In this study, a B cell epitope KNYIDKQLLPIVNKQS from the RSV F protein, known to be the target of neutralizing antibody, was chosen for vaccine development. The proposed vaccine strategy utilizes the lipid core peptide (LCP) delivery system with self-adjuvanting properties 3, 4 in conjunction with the B-cell peptide epitope and Pan DR (PADRE) as a T helper epitope (Figure 1). The vaccine candidates were designed, synthesized and their purity and identity confirmed by RP-HPLC and ESI-MS, respectively. The secondary structure analysis and potential of specific antibodies to recognize the synthetic vaccine candidates were studied. LCP delivery system as well as the coil-promoting sequence from yeast GCN4 protein was required to generate the native (desired) helical confirmation of the epitope. CD results and ELISA data indicated that candidates with helical confirmation could bind to specific antibodies. In addition, dynamic light scattering (DLS) and TEM showed that the GCN4 construct formed small nanoparticles which are expected to induce strong immune responses.

Figure 1: Schematic presentation of LCP construct.

Saranya graduated with Bachelor of Technology (Biotechnology) from Sathyabama University, India in 2009. She has obtained a Master’s degree in Biotechnology from the University of Queensland, Australia. Currently, she is pursuing her PhD under the supervision of Prof. Istvan Toth and Dr. Mariusz Skwarczynski at the University of Queensland

Group A streptococcus (GAS) infections result in a number of human diseases, including pharyngitis, rheumatic fever and rheumatic heart disease. There is no vaccine available on the market to prevent GAS. We designed and synthesized a self-adjuvanting lipopeptide (LP) GAS vaccine constructs. Each lipopeptide was composed of GAS B-cell epitopes (J14, 88-30 or their combination), an universal CD4+ T-cell helper epitope (P25) and an immunostimulant lipid moiety. The lipopeptides were synthesized using Boc-solid phase peptide synthesis (SPPS) and formulated into nanoparticles. The immunogenicity of the nanoparticles was tested in mice and antibody titres were analyzed using ELISA. Systemic IgG antibody response was elicited in outbred Swiss mice after intranasal immunization. 88/30 specific IgG response was higher in the construct containing both 88/30 and J14 epitope (LP-88/30-J14) than LP containing only 88/30 epitope (LP-88/30). All the compounds (and their formulation) were characterized with the help of dynamic light scattering (DLS), circular dichroism (CD) and transmission electron microscopy (TEM). LP-88/30-J14 formed nanoparticles of smaller size (9 nm) than LP-88/30 (50-100 nm) while LP-J14 particle size was 5 nm. The immune responses against vaccine candidates were size dependent, with the smaller particles eliciting higher antibody titers. Thus, this study showed that the choice of epitopes influenced both the particle size and the immune response against LPs.

Guangzu Zhao obtained his Master degree of Molecular Biology from The University of Queensland. He is currently in the second year of his PhD study under the supervision of Prof. Istvan Toth and Dr. Mariusz Skwarczynski. His research mainly focus on self-assemble nanoparticle vaccine.

The use of traditional vaccines based on whole microorganisms is often associated with the serious drawbacks such as allergic responses or difficulties in pathogen cultivation. These obstacles can be overcome by peptide-based vaccines. However, as peptide itself has almost no immunogenicity, adjuvant that could evoke and enhance the immune response against a supplied antigen is necessary for peptide subunit vaccine. Currently, the high toxicity and low immunogenicity of current existed adjuvants are the biggest obstacle blocked the application of peptide subunit vaccine. In this project, a novel self-adjuvanting system for peptide subunit vaccine was proposed. This system applied hydrophobic amino acids sequence to which was expected to induce self-assembly of vaccine candidates into nanoparticles. As nanoparticles are known to elicit immune response, this system should find application in vaccine development. GAS B-cell epitope J14 was used in this project to evaluate the efficiency of this system, and T-helper epitope PADRE was also applied. J14, PADRE and poly phenylalanine sequence were linked covalently through the lysine moiety to product the vaccine candidates. Two approaches were attempted to obtain the desired compound. The segment condensation based synthesis approach was unsuccessful due to poor solubility of poly phenylalanine unit in examined solvents. Stepwise synthesis approach was conducted afterwards. The whole desired compounds were synthesized successfully. Formation nanoparticles (10-70nm) were confirmed by dynamic light scattering (DLS) and transmission electron microscopy (TEM). The efficacy of particles to induce immune responses will be analysed in mice model in near future.

Wei Dai obtained a Bachelor of Biomedical Science (Biochemistry and Genetics) from the Adelaide University, Australia in 2011. After that, she obtained her Master of Biotechnology from the University of Queensland, Australia in 2013. She is currently a PhD student in the University of Queensland under the supervision of Prof. Istvan Toth and Dr. Peter M Moyle at the University of Queensland.

Gene Therapy is a technique utilized to treat diseases caused by missing, defective or overexpressing genes. Although viral vectors transfect cells efficiently, risks associated with their use limit their clinical applications1,2. Nonviral delivery systems are safer, easier to manufacture, more versatile and cost effective3. Previously studied gene delivery systems can successfully get through the cell membrane and release into cytoplasm4. However, the translocation onto the nuclear membrane and enter into the nucleus become a major challenge for gene delivery. In the current research, a library of multicomponent gnonviral gene delivery system incorporating a cationic Dendron (DEN), a cell penetrating peptide (TAT), an endosomal-disrupting peptide (GALA) and nuclear localization species (NLS) were produced (Figure 1). Four NLSs (SV40 T-antigen, nucleoplasmin, Dexamethasone and Retinoic acid) were selected to identify which one has the most efficient nuclear localization activity, and therefore is the best optimize gene delivery system.
Peptide based components were synthesized by standard solid phase peptide synthesis, and GALA was conjugated to the system by ‘click chemistry’. After finishing synthesis, these four different delivery systems will be tested and compared for DNA condensation ability, cellular uptake, and transfection efficiency. Delivery system without NLSs will be used as a negative control in the biological experiments.

Figure 1: Schematic presentation of multicomponent non-viral gene delivery system construct.

Fazren Azmi joined the Toth group as a PhD student in October 2013. Prior to this, he received his Bachelor's degree in Biotechnology with a major in drug design and development (Hons) from University of Queensland, Australia. Subsequently he was appointed as a research-lecturer fellow at the Faculty of Pharmacy, National University of Malaysia. His PhD focuses on the development of immunoadjuvants for peptide vaccine. Fazren is a recipient of the Malaysian international scholarship awarded to outstanding research higher degree students.

Peptides are of great interest to be used as vaccine antigens due to their safety, ease of manufacturing and specificity in generating immune response. There have been massive discoveries of peptide antigens over the past decade. However, peptides alone are poorly immunogenic, which demand co-administration with strong adjuvant to enhance their immunogenicity. Recently, fibril-forming peptides such as Q11 and lipoamino acid-based carrier have been identified to induce substantial immune responses when covalently linked to peptide epitope. In this study, we have incorporated either Q11 or lipoamino acids to a peptide epitope (J14) derived from M protein of Group A Streptococcus to develop self-adjuvanting vaccines. J14, Q11 and lipoamino acids were also conjugated together in a single vaccine construct in an attempt to evaluate the synergy effect of combining multiple adjuvants. Physicochemical characterization demonstrated that the vaccine constructs folded differently and self-assembled into nanoparticles. Significantly, only vaccine constructs containing double copies of lipoamino acids (regardless in conjugation with Q11 or not) were capable to induce significant dendritic cells uptake and subsequent J14-specific antibody responses in non-sizes dependent manners. Q11 had minimal impact in enhancing the immunogenicity of J14 even when it was used in combination with lipoamino acids. These findings highlight the impact of lipoamino acids moiety as a promising immunostimulant carrier and its number of attachment to peptide epitope was found to have a profound effect on the vaccine immunogenicity.

Background: Tuberculosis (TB) infection represents a global health problem and a great risk to Health Care Workers (HCWs). Identifying individuals, particularly HCWs with latent tuberculosis infection (LTBI) will support TB control through chemoprophylaxis and prevent cross-infection
Objectives: This study aimed to identify prevalence of Latent TB among a two-year new hires of HCWs in 4 major tertiary care hospitals in Riyadh, Saudi Arabia.
Material and Methods: 2650 recently (2-years) hired HCWs were surveyed for latent TB using Tuberculin Skin Test (TST). Data was collected from January 2008 to December 2009. Induration due to TST equal to or more than 10 mm. within 48-72 hours was considered positive. The results of TST were correlated with other variables such as age group, gender, job category, country of origin.
Results: As an overall rate, 291(11%) out of 2650 were positive for TST, with the highest significant positive rates among physicians (14.9%) and nurses (12.9%) compared to students as a reference group.

Introduction: Immunization against human chorionic gonadotropin (hCG) prevents pregnancy in sexually active women of proven fertility, as shown by previous Phase II efficacy trials. In order to make the vaccine consistent in its linkage to the carrier, we developed a recombinant vaccine linking hCGβ to LTB, B subunit of heat labile enterotoxin of E. coli which is a potent mucosal adjuvant. The hCGβ-LTB vaccine was fairly immunogenic in mice of different genetic strains. Since a vaccine for control of fertility should ideally be effective in every recipient and be potent enough to generate above protective threshold antibody titres to prevent pregnancy, it was decided to investigate if prime-boost approach employing a combination of anti-hCG DNA and protein vaccines, can enhance the immune response.
Methodology: hCGβ-LTB protein vaccine was made and purified using yeast Pichia pastoris pPIC9k/GS115 host-vector system. DNA version of the vaccine was prepared by incorporating the gene encoding hCGβ-LTB in eukaryotic plasmid VR1020 (DJ). Mycobacterium indicus pranii (MIP) was used as an immuno-modulator. Female inbred Balb C mice received 100 µg of DNA vaccine in saline along with 5x106 cells of MIP/animal/dose route twice fortnightly followed by 2 µg of alum adsorbed hCGβ-LTB along with 5x106 cells of MIP by intramuscular route. Second group of mice was immunized by only protein version of the vaccine along with MIP.
Results and Conclusions: Immunization with the DNA form of the recombinant hCGβ-LTB vaccine twice at fortnightly interval followed by the proteinic form of the vaccine induced distinctly higher antibody response than the proteinic vaccine alone. DNA is not only cheaper to make, it is thermostable and does not require cold chain. Hence the employment of DNA for primary immunization is expected to reduce the cost besides the benefit of enhancing antibody response.

Objective: To assess the nature of occupational exposures occurring to students and to assess the rate of reporting of incidents.
Methods: A self-administrated questionnaire was circulated to third, fourth and final year undergraduate dental students in Dental College of King Saud University, Riyadh, Kingdom of Saudi Arabia from April 2004 to May 2004. The questionnaire was consisted of 4 domains: personal details, hepatitis B vaccination, occupational exposures and reporting of the exposure incident.
Results: Significantly more incidents occurred among final year students than third year students. Through the students there was no correlation between ages, gender, and dominate hand; however, more exposures occurred in female students. A significant decrease in exposures (p<0.05) occurred when an assistant was employed. Some students with one or more exposures during their training were percutaneous injuries predominated. Seventy-seven (28.9%) students stated that they have not been vaccinated against hepatitis B virus and that was noticed among male students.
Conclusion: Dental teaching colleges face with the unique challenge of protecting the student and populations against blood-borne infections. Efforts must go beyond teaching of universal precaution, which should include safer products and clinical procedures that can reduce the risks associated with occupational exposures.

Worldwide rabies in dogs is the source of 99% of human infections. This makes dogs potential reservoir and transmitters to human being. Pre and post-exposure prophylaxis are means of protection against the rabies virus. The main objective of this experiment was to determine the safety and immunogenicity of Vero cell culture based rabies vaccine “ETHIORAB” manufactured by Ethiopian Health and Nutrition Research Institute, Vaccine and Diagnostic Production Directorate. The obtained vaccine was tested on mice and satisfactory safety results were observed. Random Clinical Trial (RCT) design was used for this experiment. Twelve experimental dogs from local breed were duly conditioned during a quarantine period and assigned to two groups randomly. Group I (cases) were vaccinated subcutaneously with 1 ml of ETHIORAB. Dogs in group II served as non-vaccinated controls. To evaluate the titer of the rabies virus neutralizing antibodies (VNA), sera were analyzed by Fluorescent Antibody Virus Neutralization (FAVN) test. Serum neutralizing antibody geomtric mean titers (GMT) to rabies virus was determined at days 7, 15, 21, 30, 60 and 90. Geomtric mean titers were equal to 1.55, 1.73, 2.02, 3.45, 3.57 and 3.17 IU/ml respectively. All dogs showed VNA titers higher than the 0.5 IU/ml mandated WHO recommended threshold. This study indicated ETHIORAB rabies vaccine manufactured in Ethiopia is safe and immunogenic.

Abstract [Track 1-1 Drug interactions and pharmacological compatibilities] Metformin interaction with popular Indian herbal remedies, Boswellia serrata (Sallaki) and Withania somnifera (Ashwagandha) – PK/PD approach with insulin resistant rodent model Arun T S, Rekha R Shenoy, C Mallikarjuna Rao and N Gopalan Kutty Department of Pharmacology, Manipal College of Pharmaceutical Sciences, Manipal University The general perception of herbal medicines as a safer alternative to allopathic drugs often leads to self-medication. Concomitant usage of herbal remedies (by chance or choice) with prescription medicines can lead to un-intended interactions modifying pharmacological properties of either drug, which could lead to potential treatment failure.
We designed a study, where metformin [MET], an extensively prescribed anti-diabetic medication was evaluated with commonly used Indian herbal remedies, Sallaki (SAL) and Ashwagandha (ASH). Pharmacokinetic and pharmacodynamics profiles were determined under normal and insulin resistance conditions in rodents. A sensitive bioanalytical method was developed and validated in-house for the determination of metformin. Pharmacokinetic and pharmacodynamic evaluations were performed by using Winnonlin (v 5.3). The evaluations included estimation of Cmax, tmax, AUC0-t, AUC0-∞, Vd, Cl, t1/2 and Kel by non-compartmental pharmacokinetic analysis and estimation of plasma glucose concentrations at the scheduled time along with PK sampling. The data assessment involved plotting of response (plasma glucose) vs time and subsequent group wise comparison of the obtained data in both actual and percent-normalized form. The analysis of response vs time data was performed by non-compartmental pharmacodynamics analysis WinNonlin version 5.3.
The PK-PD study demonstrated that increased obesity and associated pathological changes led to increased systemic exposure of metformin. In the absence of herbs, MET glucose profile was stable with a narrow range of plasma glucose variations. ASH does not affect metformin pharmacokinetics. The Cmax was reduced by 37% and AUC by 47%. Administration of SAL reduces the bioavailability of metformin. In presence of ongoing SAL treatment, higher doses of MET could be required to produce required effect.
This study assumes greater importance considering the fact that the chosen herbal remedies and the drug are used for the treatment of chronic conditions. Not many interactions are reported for metformin owing to its physico-chemical and metabolic characteristics. Our findings demonstrate that concomitant usage of metformin with sallaki may adversely affect the outcome or may result in needless increase in dosage.

Anoop Kumar has submitted his PhD at the age of 27 years from University of delhi, Delhi, India. Thesis title “Development, Characterization & Immunogenicity of a novel HPV-16 Vaccine construct” targeting the major health problem in low resource countries like India.

In India, cervical cancer is the most frequent gynecological cancer and approximately 432.20 million Indian women above 15 years of age are at risk of developing cervical cancer. India contributes about 1/3rd of the global cervical cancer deaths, with estimates of 132,000 new cases diagnosed and 74,000 deaths annually. The persistent infection with high risk-HPV (HR-HPV) is the major cause of cervical cancer, worldwide. Two developed prophylactic HPV vaccine (Gardasil and Cerverix) provide protection against HPV induced cervical malignancy. However, vaccines having therapeutic values are of utmost priority. Till date, most of HPV therapeutic vaccines are focused on two major HPV oncoproteins (E6/E7). HPV-E5 which acts by altering the activity of cellular proteins, mainly growth factor pathways emerges as a new therapeutic target. In the present study, we predicted the epitopes of HPV16-E5 to identify the candidate B-cell and T-cell epitopes, which can be used for HPV immunotherapy. We identified that epitope SAFRCFIVYIIFVY is the most potent peptide for HLA-A*11:01 having percentile value of 0.5 and immunogenicity score of 0.69558. For MHC-II, the epitopes IPLFLIHTHARFLIT for HLA-DRB1*14:01 alleles have the lowest IC50 value (18.13nM). The identification of structural feature and immunogenic epitopes provides the best information for development of drugs or vaccine. In conclusion, as the expression of E5 protein was detected in the early phase of the HPV infection, there may be an opportunity to target HPV-E5 that would help in the prevention and progression of the precancerous lesion to invasive carcinomas.

Mr. Rajesh Jain, M Pharm in Pharmaceutics from Mumbai University, is currently persuing PhD in Pharmaceutics from the Bombay College of Pharmacy, Mumbai, INDIA. Research Experience: 5 years; Master research work involved development of delivery sytem for vaccine and evaluation of same in suitable animal model. PhD research work is development of targeted inhalation drug delivery sytems.; Industry Experience- - 1.5 yrs after completion of M Pharm; Publications- -5; National Presentations- 10; Book chapters- 1.

Dental caries is a painful, morbid condition, manifested mainly in children, as localized destruction of susceptible dental hard tissues by acidic by-products from bacterial fermentation of dietary carbohydrates. Treatment is painful, time consuming, expensive, involves drilling and filling, Several microorganisms have been identified in carious lesions; the members of mutans streptococci are the main causative agents. A preventive approach by interfering with initial S. mutans colonization, which can be facilitated by a vaccine, administered by mucosal route, would be more useful. A Panel on Caries Vaccine Sponsored by The National Institute of Dental and Craniofacial Research (NIDCR), NIH has identified several antigenic targets on S.mutans for dental caries vaccine. The aim of this investigation was to study the feasibility of achieving mucosal immunization against dental caries using antigens from Streptococcus mutans, for intranasal and sublingual administration, which would be more acceptable to children, than an injection. In the first step, key antigens (NIDCR) from S. mutans were extracted, purified and characterized. Further, in vivo efficacy studies in Wistar rats were carried out to assess comparative antibody titers (IgG & IgA) in serum, saliva, nasal lavage and BAL, on administration of these antigens via three routes- intramuscular (IM), intranasal (IN) and sublingual (SL) ; antibody titres being determined by indirect ELISA technique. Increased induction and persistence of humoral immune responses (IgG in serum) in both IN and SL group, compared to the invasive IM route was evident. Also, significant antibody titers (IgA) in saliva, nasal lavage and BAL, in comparison to IM routes were seen. (Unpaired t test, p<0.05) In conclusion, mucosal immunization by intranasal and sublingual routes for the S. mutans antigens indicates the potential of the mucosal vaccine delivery approach for developing a needle-free vaccine for prevention of dental caries in children.