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The mycobacterial glycolipid glucose monomycolate induces a memory T cell response comparable to a model protein antigen and no B cell response upon experimental vaccination of cattle.

Nguyen TK, Koets AP, Santema WJ, van Eden W, Rutten VP, Van Rhijn I - Vaccine (2009)

Bottom Line: Glycolipids are presented to T cells by human group 1 CD1 proteins, but are not used as subunit vaccines yet.Also, KLH induced strong antibody responses whereas GMM did not.These data suggest that non-overlapping T cell populations are targeted and demonstrate the potential of glycolipids as a special class of subunit vaccine candidates.

View Article: PubMed Central - PubMed

Affiliation: Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, Utrecht, The Netherlands.

ABSTRACT
Glycolipids are presented to T cells by human group 1 CD1 proteins, but are not used as subunit vaccines yet. Experimental immunizations with pure mycobacterial glucose monomycolate (GMM) and keyhole limpet haemocyanin (KLH) in cattle, a species which, unlike mice, expresses group 1 CD1, showed that GMM was equally efficient as KLH in generating T cell responses in blood, but not in the draining lymph node. Also, KLH induced strong antibody responses whereas GMM did not. These data suggest that non-overlapping T cell populations are targeted and demonstrate the potential of glycolipids as a special class of subunit vaccine candidates.

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T cell responses against GMM and KLH, induced by immunization. (A) Freshly isolated PBMC of GMM/KLH-immunized animals and adjuvant only immunized animals were stimulated with GMM extracted from N. farcinica and M. phlei. The cross-reactive T cell stimulation shown is measured by stimulating the T cells with the GMM that was not used for immunization. (B) In the same assay, the T cell response against the protein antigen KLH was determined. The T cell stimulation index was calculated by dividing the [3H]thymidine incorporation of the antigen-stimulated PBMC by the [3H]thymidine incorporation of PBMC cultured in medium without lipids (Table 1) after a 8-h [3H]thymidine pulse performed after a three-day incubation. Each symbol represents the mean stimulation index (±standard deviation) of eight independent experiments performed post-immunization on one animal. Black symbols represent the GMM/KLH-immunized animals and white symbols represent adjuvant only immunized animals. (C) PBMCs of a GMM/KLH-immunized animal were incubated with GMM or medium only in the presence of the cross-reactive anti-human CD1b monoclonal antibody BCD1b.3, or the isotype control P3. Bars represent the mean CPM of triplicate wells (±standard deviation).
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fig1: T cell responses against GMM and KLH, induced by immunization. (A) Freshly isolated PBMC of GMM/KLH-immunized animals and adjuvant only immunized animals were stimulated with GMM extracted from N. farcinica and M. phlei. The cross-reactive T cell stimulation shown is measured by stimulating the T cells with the GMM that was not used for immunization. (B) In the same assay, the T cell response against the protein antigen KLH was determined. The T cell stimulation index was calculated by dividing the [3H]thymidine incorporation of the antigen-stimulated PBMC by the [3H]thymidine incorporation of PBMC cultured in medium without lipids (Table 1) after a 8-h [3H]thymidine pulse performed after a three-day incubation. Each symbol represents the mean stimulation index (±standard deviation) of eight independent experiments performed post-immunization on one animal. Black symbols represent the GMM/KLH-immunized animals and white symbols represent adjuvant only immunized animals. (C) PBMCs of a GMM/KLH-immunized animal were incubated with GMM or medium only in the presence of the cross-reactive anti-human CD1b monoclonal antibody BCD1b.3, or the isotype control P3. Bars represent the mean CPM of triplicate wells (±standard deviation).

Mentions: To study the efficiency of T cell priming upon immunization with a glycolipid antigen as compared with a known immunogenic model protein, cattle (n = 7) were immunized with GMM and KLH in DDA. To reduce the effects of inter-individual differences, while preventing any possible interaction of the two antigens when mixed, the animals received the two antigens separately, in the right and the left shoulder, on the same day. Another group of animals (n = 5) of the same age and sex, housed together with the GMM/KLH immunized group, was immunized with adjuvant only and served as a control group. Before the first immunization and from 1 week till 4.5 months after the second immunization, animals were tested every other week for T cell reactivity against GMM and KLH. The GMM specific proliferative T cell response that was detected in freshly isolated PBMC after immunization with GMM/KLH was significantly higher than in adjuvant only immunized animals (P < 0.001) (Fig. 1A). A similar pattern was observed for KLH when GMM/KLH immunized and adjuvant only immunized groups were compared (Fig. 1B). Of note, the strength of the response against KLH and against GMM was comparable. Before immunization there were no significant differences between the groups (not shown).


The mycobacterial glycolipid glucose monomycolate induces a memory T cell response comparable to a model protein antigen and no B cell response upon experimental vaccination of cattle.

Nguyen TK, Koets AP, Santema WJ, van Eden W, Rutten VP, Van Rhijn I - Vaccine (2009)

T cell responses against GMM and KLH, induced by immunization. (A) Freshly isolated PBMC of GMM/KLH-immunized animals and adjuvant only immunized animals were stimulated with GMM extracted from N. farcinica and M. phlei. The cross-reactive T cell stimulation shown is measured by stimulating the T cells with the GMM that was not used for immunization. (B) In the same assay, the T cell response against the protein antigen KLH was determined. The T cell stimulation index was calculated by dividing the [3H]thymidine incorporation of the antigen-stimulated PBMC by the [3H]thymidine incorporation of PBMC cultured in medium without lipids (Table 1) after a 8-h [3H]thymidine pulse performed after a three-day incubation. Each symbol represents the mean stimulation index (±standard deviation) of eight independent experiments performed post-immunization on one animal. Black symbols represent the GMM/KLH-immunized animals and white symbols represent adjuvant only immunized animals. (C) PBMCs of a GMM/KLH-immunized animal were incubated with GMM or medium only in the presence of the cross-reactive anti-human CD1b monoclonal antibody BCD1b.3, or the isotype control P3. Bars represent the mean CPM of triplicate wells (±standard deviation).
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fig1: T cell responses against GMM and KLH, induced by immunization. (A) Freshly isolated PBMC of GMM/KLH-immunized animals and adjuvant only immunized animals were stimulated with GMM extracted from N. farcinica and M. phlei. The cross-reactive T cell stimulation shown is measured by stimulating the T cells with the GMM that was not used for immunization. (B) In the same assay, the T cell response against the protein antigen KLH was determined. The T cell stimulation index was calculated by dividing the [3H]thymidine incorporation of the antigen-stimulated PBMC by the [3H]thymidine incorporation of PBMC cultured in medium without lipids (Table 1) after a 8-h [3H]thymidine pulse performed after a three-day incubation. Each symbol represents the mean stimulation index (±standard deviation) of eight independent experiments performed post-immunization on one animal. Black symbols represent the GMM/KLH-immunized animals and white symbols represent adjuvant only immunized animals. (C) PBMCs of a GMM/KLH-immunized animal were incubated with GMM or medium only in the presence of the cross-reactive anti-human CD1b monoclonal antibody BCD1b.3, or the isotype control P3. Bars represent the mean CPM of triplicate wells (±standard deviation).
Mentions: To study the efficiency of T cell priming upon immunization with a glycolipid antigen as compared with a known immunogenic model protein, cattle (n = 7) were immunized with GMM and KLH in DDA. To reduce the effects of inter-individual differences, while preventing any possible interaction of the two antigens when mixed, the animals received the two antigens separately, in the right and the left shoulder, on the same day. Another group of animals (n = 5) of the same age and sex, housed together with the GMM/KLH immunized group, was immunized with adjuvant only and served as a control group. Before the first immunization and from 1 week till 4.5 months after the second immunization, animals were tested every other week for T cell reactivity against GMM and KLH. The GMM specific proliferative T cell response that was detected in freshly isolated PBMC after immunization with GMM/KLH was significantly higher than in adjuvant only immunized animals (P < 0.001) (Fig. 1A). A similar pattern was observed for KLH when GMM/KLH immunized and adjuvant only immunized groups were compared (Fig. 1B). Of note, the strength of the response against KLH and against GMM was comparable. Before immunization there were no significant differences between the groups (not shown).

Bottom Line: Glycolipids are presented to T cells by human group 1 CD1 proteins, but are not used as subunit vaccines yet.Also, KLH induced strong antibody responses whereas GMM did not.These data suggest that non-overlapping T cell populations are targeted and demonstrate the potential of glycolipids as a special class of subunit vaccine candidates.

View Article: PubMed Central - PubMed

Affiliation: Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, Utrecht, The Netherlands.

ABSTRACT
Glycolipids are presented to T cells by human group 1 CD1 proteins, but are not used as subunit vaccines yet. Experimental immunizations with pure mycobacterial glucose monomycolate (GMM) and keyhole limpet haemocyanin (KLH) in cattle, a species which, unlike mice, expresses group 1 CD1, showed that GMM was equally efficient as KLH in generating T cell responses in blood, but not in the draining lymph node. Also, KLH induced strong antibody responses whereas GMM did not. These data suggest that non-overlapping T cell populations are targeted and demonstrate the potential of glycolipids as a special class of subunit vaccine candidates.

Show MeSH