The potency of a modified Alum precipitated haemorrhagic septicaemia vaccine in mice, rabbit and cattle was evaluated. Active mouse protection test showed that the log of protection was 7.8. Passive mouse protection test on sera obtained from vaccinated cattle showed 67-100% protection up to four months and 25-67% protection 2 months post vaccination. Modified Alum precipitated vaccine was potent to produce immunity against active challenge with virulent Pasteurella multocida in rabbits. The antisera collected from vaccinated rabbits conferred 83-100% protection in surmised mice.

Objective(s): Many types of human papillomaviruses (HPVs) have been identified, with some leading to cancer and others to skin lesions such as anogenital warts. Studies have demonstrated an association between oncogenic HPV and cervical cancer and many researchers have focused on therapeutic vaccines development. At present, the modulatory effect of opioids on the innate and acquired immune system is characterized. Antagonists of opioid receptors such as naloxone (NLX) can contribute to the shifting Th2 response toward Th1. Herein; we studied the adjuvant activity of NLX/Alum mixture for improvement of the immunogenicity of HPV-16E7d vaccine. Materials and Methods: The mice were administered different regimens of vaccine; E7d, E7d-NLX, E7d-Alum, E7d-NLX-Alum, NLX, Alum and PBS via subcutaneous route for three times with two weeks interval. Two weeks after the last immunization, the sera were assessed for total antibody, IgG1 and IgG2a with an optimized ELISA method. The splenocytes culture supernatant was analyzed by ELISA for the presence of IL-4, IFN- and IL-17 cytokines and lymphocyte proliferation was evaluated with Brdu method. Results: Immunization of mice with HPV-16 E7d vaccine formulated in NLX/Alum mixture significantly increased lymphocyte proliferation and Th1 and Th17 cytokines responses compared to other experimental groups. Analysis of humoral immune responses revealed that administration of vaccine with NLX/Alum mixture significantly increased specific IgG responses and also isotypes compared to control groups. Conclusion: NLX/Alum mixture as an adjuvant could improve cellular and humoral immune responses and the adjuvant maybe useful for HPV vaccines model for further studies in human clinical trial.

Background: adjuvants are essential to potentiate the immune response to inoculated antigens and play a central role in vaccine development. Alum is generally used as a classic adjuvant, although it does not stimulate proper immunity, and some of the immunized subjects have low or no antibody response. Efforts have been continued to find more efficient adjuvants for better antibody responses. In the present study, the efficacy of three formulations of adjuvants, i. e. Cysteine p Guanine Oligodeoxynucleotide (CpG ODN), Alum, and Freund, in the production of monoclonal anti Hepatitis B Surface Antigen (HBsAg) antibodies was investigated. Methods: To immunize mice, regular hepatitis B vaccine containing recombinant HBsAg and Alum was used with CpG ODN or Freund adjuvants, and splenocytes of hyperimmunized mice were fused with murine myeloma Sp2/0 cells. Positive hybridomas were selected by Enzyme-Linked Immunosorbent Assay (ELISA) using HBsAg as coating antigen followed by a limited dilution process. Results: The results showed that by using all three formulations of adjuvants, monoclonal antibody (mAb) specific to HBsAg was successfully generated. It was also found that the mice immunized with (HBsAg + Alum) + CpG had the highest concentration of antibody production in serum and hybridoma supernatants as well as positive clones. Based on these findings, the addition of CpG ODN also induced a higher antibody response compared with Complete Freund’ s adjuvant (CFA). Conclusion: Results of this study showed that CpG and Freund adjuvants could be efficient partners for Alum in the immunization period of the process of monoclonal antibody production.

Cytokines have important roles in the control of bacterial and viral infections such as HIV-1. Interleukin 17 which is secreted by Th17 is one of these cytokines with a special role in controlling microbial infections. In the present study, adjuvant activity of Alum and Naloxone mixture has been studied on immune responses, especially IL-17 cytokine. Naloxone and Alum adjuvant are mixed with 10 mg of recombinant vaccine HIV-1-gag-pol-tat-env. Experimental groups, consisting of 36 inbred male Balb/c mice divided into six groups, were injected sub cutaneouslyat days 0, 14 and 28 with total volume of 200 ml.2 weeks after final injection, mouse spleens were removed in sterile conditions and used to prepare suspensions. Lymphocyte proliferation responses were evaluated with Brdu test and evaluation of cytokines IL-4, IL-17 and INF-γ were completed using ELISA kit, plus total antibody and antibody isotopes IgG1 and IgG2a using ELISA test. All results show that the mixture of Alum with Naloxone increased cellular immune parameters and specially raised interleukin 17 which illustrated a significant difference with other groups. It seems that Alum and Naloxone mixture could control viral infections by affecting the Th17 pathway in which IL17 cytokine has a critical role.

Purpose: The introduction of novel adjuvants is an important step in attempts to develop a safe and more efficient vaccine. The present study was performed to determine whether the use of a mixed beta-adrenergic receptor antagonist propranolol (PRP) and Aluminum (Alum), as an adjuvant, have efficacy for Toxoplasma gondii vaccine to induce protective immunity in a mouse model. Methods: Female BALB/c mice divided into five groups were immunized with excretorysecretory antigens (ESA) vaccine, Alum-ESA vaccine, PRP-ESA vaccine, and Alum-PRP ESA vaccine, as well as with phosphate buffered saline (PBS), as a negative control group. The immune responses were evaluated by lymphocyte proliferation assay for measuring delayedtype hypersensitivity (DTH) response and by cytokine assay for evaluating IFN-γ and IL-5 levels. The survival rate of mice in all groups was assessed during a three-week monitoring period after an intraperitoneal challenge with T. gondii tachyzoites. Results: The results showed that mice immunized with PRP, as an adjuvant, could secret a higher level of IFN-γ , which was significant in comparison to other groups. However, mice vaccinated with Alum-precipitated ESA antigen had ability to produce an elevated level of IL-5 compared to other mouse groups (P≤ 0. 05). Moreover, Alum-PRP co-administration together with ESA vaccine resulted in the longer survival of mice. Conclusion: The findings of this study revealed that the combination of Alum-PRP adjuvants and ESA vaccine of T. gondii elicits both humoral and cellular immune responses, which are comparable to either Alum or PRP alone.

Objective(s): SARS-CoV-2, emerging as a major threat to public health, has to be controlled through vaccination. Naloxone (NLX), an opioid receptor antagonist, demonstrated its adjuvant activity for microbial vaccines. In this study, inactivated SARS-CoV-2 was developed in the Alum/NLX adjuvant to increase the potency of the inactivated SARS-CoV-2 vaccine. Materials and Methods: BALB/c mice were immunized on days 0 and 14 with inactivated SARS-CoV-2-Alum,-Alum + NLX 3 mg/kg,-Alum + NLX 10 mg/kg, and-Freund adjuvant, as well as PBS. IFN-γ, and IL-4 cytokines and Granzyme-B release were assessed with ELISA. In addition, specific total IgG, IgG1/IgG2a isotypes, and ratio as well as anti-RBD IgG responses were assessed with an optimized ELISA. Results: SARS-CoV-2-Alum-NLX10 group showed a significant increase in the IFN-γ, cytokine response versus SARS-CoV-2-Alum, SARS-CoV-2-Alum-NLX3, and PBS groups. The SARS-CoV-2-Alum-NLX3 group exhibited a significant decrease in IL-4 cytokine versus SARS-CoV-2-Alum. The mice immunized with SARS-CoV-2-Alum-NLX10 showed a significant increase in CTL activity versus SARS-CoV-2-Alum and PBS. In addition, mice immunized with SARS-CoV-2-Alum-NLX3, SARS-CoV-2-Alum-NLX10 and SARS-CoV-2-Freund demonstrated an increase in IgG response, as compared with SARS-CoV-2-Alum and PBS group. Furthermore, all formulations of SARS-CoV-2 vaccines could induce both IgG1 and IgG2a isotypes. But, the IgG2a/IgG1 ratio in SARS-CoV-2-Freund and SARS-CoV-2-Alum-NLX10 revealed an increase as compared with that of the SARS-CoV-2-Alum group. Anti-RBD IgG response in the SARS-CoV-2-Alum-NLX10 group showed a significant increase as compared with the Alum-based vaccine. Conclusion: Formulation of inactivated SARS-CoV-2 virus in NLX/Alum adjuvant improved the potency of humoral and, especially, cellular responses.