Clin Exp Vaccine Res.  2015 Jan;4(1):68-74. 10.7774/cevr.2015.4.1.68.

Cellular immune response following pre-exposure and postexposure rabies vaccination by intradermal and intramuscular routes

• Venkataswamy MM
• Madhusudana SN
• Sanyal SS
• Taj S
• Belludi AY
• Mani RS
• Hazra N

• KMID: 2049109
• DOI: http://doi.org/10.7774/cevr.2015.4.1.68

Abstract

PURPOSE
Immunization against rabies in humans induces protective neutralizing antibodies; however, the induction of type 1 or type 2 cytokine mediated cellular immune responses following rabies vaccination is not understood. Hence, the present study investigated cellular cytokine responses in vaccinated individuals.
MATERIALS AND METHODS
The study groups included healthy rabies antigen naive controls (n=10), individuals who received intradermal primary (n=10) or booster pre-exposure vaccination (n=20) and subjects who received postexposure rabies vaccination either by intradermal (n=18) or intramuscular (n=20) routes. The antigen specific cellular responses were analyzed by stimulating peripheral blood mononuclear cells with a rabies vaccine antigen in the interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) enzyme-linked immunospot (ELISpot) assay. These responses were compared to the rabies virus neutralizing antibody (RVNA) titers that were measured by rapid fluorescent focus inhibition test.
RESULTS
We observed that cellular and humoral immune responses to primary intradermal rabies vaccination could be greatly enhanced by a booster vaccine; and both type 1 and type 2 cytokine responses were significantly elevated. The magnitude of type 1 and type 2 cytokine responses did not differ significantly among the intramuscular and intradermal routes of postexposure vaccination. The number of cells producing IFN-gamma and IL-4 correlated significantly with the levels of RVNA.
CONCLUSION
Both type 1 and type 2 cellular cytokine responses are strongly induced after rabies vaccination and directly correlate with levels of RVNA titers. The neutralizing antibody as well as the type 1 and type 2 cytokine responses may be important for vaccine induced protective responses against rabies.

Keyword

Rabies; Rabies prophylaxis; Rabies vaccines; IFN gamma; IL4; Immune response

MeSH Terms

Antibodies, Neutralizing
Humans
Immunity, Cellular*
Immunity, Humoral
Immunization
Interferon-gamma
Interleukin-4
Rabies Vaccines
Rabies virus
Rabies*
Vaccination*
Antibodies, Neutralizing
Interferon-gamma
Interleukin-4
Rabies Vaccines

Figure

• 

  Fig. 1 Enzyme-linked immunospot assay for rabies specific interferon-γ (IFN-γ) (A) and interleukin-4 (IL-4) (B) producing T cells in peripheral blood mononuclear cells (PBMCs), seven days after pre-exposure primary intradermal (ID) vaccination and after ID booster dose. (C) Rapid fluorescent focus inhibition test assay results for rabies virus neutralizing antibody (RVNA) titers in serum, seven days after primary ID vaccination and after ID booster dose. SFC, spot-forming cell. **p < 0.005 and ***p < 0.001 (Mann Whitney test). Pearson's correlation co-efficient analysis of antibody titres and cellular cytokine responses between naive, ID pre-exposure prime and boost groups. (D) RVNA versus IFN-γ. (E) RVNA versus IL-4.

• 

  Fig. 2 Representative image for enzyme-linked immunospot (ELISpot) responses from a healthy control, individual who received pre-exposure primary intradermal (ID) vaccination six months earlier and an individual who received booster ID vaccine at six months. The top row of wells represent the unstimulated peripheral blood mononuclear cell (PBMC) control wells while the bottom row represent the rabies antigen stimulated wells. The results from interferon-γ (A) and interleukin-4 (B) ELISpot assays are shown.

• 

  Fig. 3 Enzyme-linked immunospot assay for rabies specific interferon-γ (IFN-γ) (A) and interleukin-4 (IL-4) (B) producing T cells in peripheral blood mononuclear cells (PBMCs), seven days after postexposure primary intradermal (ID) and intramuscular (IM) vaccination. (C) Rapid fluorescent focus inhibition test assay for rabies virus neutralizing antibody (RVNA) titers in serum, seven days after postexposure primary ID and IM vaccination. ***p < 0.001 (Mann Whitney test).

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