Karine SERRE, PhD
IMMUNOLOGIST
Latest Posts
21/01/2016
Psoriasis is an autoimmune disease and NOT a chronic inflammatory disease!12/05/2015
CALL For PAPERS: “Innate T Cell Development and Functions in Immune Diseases”28/08/2014
XL Annual Meeting of the Sociedade Portuguesa de Imunologia 13-15th October 201413/04/2013
Beginnings of breaking down the differentiation program of innate IL-17-production27/01/2013
The improbable trio NKG2D – γδ T cells – IgE!!Influence of antigen-specific CD8 T cells on the B cell responses to alum-precipitated antigen
IFN-g produced by CD8 T cells induces class switching in B cells in responses to alum-protein vaccine
Alum-protein vaccines are widely used for their ability to elicit long-lasting neutralizing antibody responses that prevent bacterial exotoxins and viruses entering cells. Typically these vaccines induce Th2-dependent antibody responses characterized by the differentiation of IL-4-producing CD4 T cells and immunoglobulin class switching to IgG1 (1).
Thus, CD4 cells in response to these antigens produce IL-4 and IL-13. However, responding CD8 T cells show no signs of T cytotoxic 2 (Tc2) commitment (2). This contrasts with the reported ability of naïve CD8 T cells, when stimulated in presence of IL-4 in vitro, to differentiate into IL-4-producing Tc2 (3-5). In addition IL-13 and IL-5-producing Tc2 cells have been reported in a mouse model of airway inflammation (6, 7) and Tc2 clones have been derived from human blood (8, 9). By contrast, CD8 T cells have been found to respond to alum-proteins by proliferating extensively and producing IFN-g, a key Th1 cytokine (2).
Based on these findings, we questioned whether adoptive transfer of antigen-specific CD8 T cells would affect the characteristic Th2 response of CD4 T cells and the nature of the antibody response to alum-proteins. To this end wild type mice received transgenic ovalbumin (OVA)-specific CD4 (OTII) or CD8 (OTI) T cells or both of these, and were immunized with alum-precipitated OVA (alumOVA) in both footpads.
In these settings the OTI and OTII cell response is presented here. In the present page I will show the influence of the OTI response on the endogenous B cell response.
This work was done in collaboration with Dr. Elodie Mohr from the MRC centre for immune regulation at the University of Birmingham, UK (Dr. Mohr current address is now: Instituto Gulbenkian de Ciência, Oeiras, Portugal).
CD8 OTI cells diversify the B cell switching pattern in the response to alumOVA
In chimeras with OTII cells alone these CD4 T cells typically produce Th2 cytokines in response to alumOVA and provide help for both follicular and extrafollicular B cells responses with switching to IgG1 (1, 10, 11). 
The endogenous CD4 T cell help has only a limiting effect on the rate of development of this antibody response (Fig. 1A). Seven days after immunization with alumOVA, the number of CD138+B220Int antibody forming cells (AFC) produced in LN in chimeras constructed with OTII cell transfer is 400 times that in WT mice. As expected the OTII-assisted response predominantly induces switching to IgG1 (Fig. 1B). Somewhat surprisingly CD8 T cells also provide help for AFC production, for chimeras constructed with OTI cells had some 8 fold more AFC than WT controls 7 days after immunization. The co-transfer of CD8 (OTI) with CD4 (OTII) cells did not appreciably alter the number of AFC produced compared to the number of AFC generated in OTII cell chimeras. Nevertheless, the phenotype of the AFC in the mixed chimeras was strikingly altered. In the mixed chimeras switching to IgG1 was greatly reduced and many AFC had switched to IgG2a or IgG2b (Fig. 1B).
IFN-g production in mixed chimeras is in large part responsible for the loss of switching to IgG1 and induction of switching to IgG2a and IgG2b
We have previously shown that OTI cells responding to alumOVA do not acquire the properties of producing IL-4 or IL-13, but instead proliferate and a large proportion of the primed cells secrete IFN-g (see here and (2)). This is of interest, for IFN-g can suppress switching to IgG1 or IgE and promote switching to IgG2a (12-15). Consequently, we next tested whether the effect of OTI cells on switching in the mixed chimeras was attributable to the IFN-g production induced in these mice.
To do this, chimeras were constructed and immunized as before, but neutralizing antibody against IFN-γ or isotype control antibody was given at the time of transfer and 3 days after immunization. Neutralizing anti-IFN-g antibody largely reverses the effect of transferring OTI cells with OTII cells (Fig. 2). Switching to IgG1 is restored and there is significantly fewer switched IgG2a and IgG2b AFC (Fig. 2). Thus in large part of CD8 T cell-directed switching to IgG2a and IgG2b and suppression of switching to IgG1 are dependent upon the IFN-g.
Conclusions
These data show that the presence of antigen-specific CD8 T cells during T-dependent antibody responses to alum-precipitated proteins, skews B cell switching from IgG1 to IgG2a and IgG2b. IFN-g produced by the activated CD8 T cells plays a major part in re-directing the response of CD4 T cells and B cells from Th2 to Th1. At this stage of the study, when and where the influence of the activated CD8 T cells occurs is unclear.
During T-dependent responses to alum-proteins, Ig class switching takes place in the outer T zone or paracortex of the LN, or later in the germinal centers (GC) (16, 17). This switching generally requires a transient interaction between CD4 T cell CD40L and B cell CD40 (18). Activated CD8 T cells can express CD40L (19-21), but typically they are not found in the follicles ((22) and data not shown).
Consistent with this, OTI cells do not acquire follicular T helper cell features including expression of CXCR5 and PD-1, whereas a substantial number of OTII cells in the model described in this paper do (2). Thus, the IFN-g produced by responding OTI cells may either directly influence B cell switching in the outer T zone, or indirectly modify the way CD4 T cells induce B cells to switch; a combination of these may also operate. Further studies are warranted to test these hypotheses.
This model provide further insight into the way alum vaccines coordinate the activation of different actors of the adaptive immune system -the antigen-specific CD8 T cells and B cells. While aluminum adjuvants induce Th2 type of responses through Toll-like receptor (TLR) signaling-independent pathways (25-28), our data show that recruitment of antigen-specific CD8 T cells bypass the need for TLR agonists to achieve B cell switching to Th1 Ig isotypes. This mechanism may offer an option to modulate B cell responses after vaccination protocol in the presence of alum adjuvant.
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