Limits...
Bee venom processes human skin lipids for presentation by CD1a.

Bourgeois EA, Subramaniam S, Cheng TY, De Jong A, Layre E, Ly D, Salimi M, Legaspi A, Modlin RL, Salio M, Cerundolo V, Moody DB, Ogg G - J. Exp. Med. (2015)

Bottom Line: Patient studies showed that injected PLA2 generates lysophospholipids within human skin in vivo, and polyclonal T cell responses are dependent on CD1a protein and PLA2.These findings support a previously unknown skin immune response based on T cell recognition of CD1a proteins and lipid neoantigen generated in vivo by phospholipases.The findings have implications for skin barrier sensing by T cells and mechanisms underlying phospholipase-dependent inflammatory skin disease.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Rheumatology, Immunology and Allergy, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, 02114.

Show MeSH

Related in: MedlinePlus

CD1a-restricted reactivity to Apis mellifera venom PLA2 is found among polyclonal T cells from blood of healthy donors. CD3+ cells were isolated by magnetic beads from healthy donor PBMC and cultured for 12–14 d with IL-2 and irradiated K562 cells transfected with CD1a (K562 CD1a) or CD1b (K562 CD1b) or CD1c (K562 CD1c) or CD1d (K562 CD1d) or an empty vector (K562) in the presence of Apis mellifera venom PLA2. CD1 reactivity was then examined by IFN-γ ELISpot with transfected or untransfected K562 cells either in the presence or absence of Apis mellifera venom 100 ng/ml PLA2 (A, left). Data from one representative donor of three are shown. Venom PLA2 specific CD1a-restricted T cell responses were measured in 18 donors (A, right). CD1a-restricted responses were examined in the presence or absence of anti-CD1a (B, left) for donor C1098, and PLA2-neutralizing antibodies (7B, right), and manoalide (specific PLA2 inhibitor) for wasp venom in donor C559 (C, left) and Apis mellifera venom PLA2 in donor C334 (C, right). Representative results are shown from three independent experiments. (D) IFN-γ production was measured by co-incubations of PLA2-specific T cells with mDCs (left) or CD14-derived LC-like DCs (middle) in the presence or absence of anti-CD1a for donor C558. IFN-γ production was measured from co-incubations of PLA2-specific T cells and freshly isolated skin CD1a+ cells in the presence or absence of anti-CD1a or manoalide (right). Representative data from 3 (left and middle) or 2 (right) separate donors are shown. Data are mean of triplicate measurements ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4322046&req=5

fig7: CD1a-restricted reactivity to Apis mellifera venom PLA2 is found among polyclonal T cells from blood of healthy donors. CD3+ cells were isolated by magnetic beads from healthy donor PBMC and cultured for 12–14 d with IL-2 and irradiated K562 cells transfected with CD1a (K562 CD1a) or CD1b (K562 CD1b) or CD1c (K562 CD1c) or CD1d (K562 CD1d) or an empty vector (K562) in the presence of Apis mellifera venom PLA2. CD1 reactivity was then examined by IFN-γ ELISpot with transfected or untransfected K562 cells either in the presence or absence of Apis mellifera venom 100 ng/ml PLA2 (A, left). Data from one representative donor of three are shown. Venom PLA2 specific CD1a-restricted T cell responses were measured in 18 donors (A, right). CD1a-restricted responses were examined in the presence or absence of anti-CD1a (B, left) for donor C1098, and PLA2-neutralizing antibodies (7B, right), and manoalide (specific PLA2 inhibitor) for wasp venom in donor C559 (C, left) and Apis mellifera venom PLA2 in donor C334 (C, right). Representative results are shown from three independent experiments. (D) IFN-γ production was measured by co-incubations of PLA2-specific T cells with mDCs (left) or CD14-derived LC-like DCs (middle) in the presence or absence of anti-CD1a for donor C558. IFN-γ production was measured from co-incubations of PLA2-specific T cells and freshly isolated skin CD1a+ cells in the presence or absence of anti-CD1a or manoalide (right). Representative data from 3 (left and middle) or 2 (right) separate donors are shown. Data are mean of triplicate measurements ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Mentions: Having identified PLA2 as sufficient to generate CD1a-presented antigens in vitro using T cell clones, we next sought to determine if bee venom PLA2 is sufficient to activate polyclonal T cells ex vivo in cellular assays (Fig. 7). As with whole venoms (Fig. 1), we detected higher IFN-γ–producing cells with the addition of 100 ng/ml bee venom PLA2 but only in the presence of CD1a and not other CD1 isoforms (Fig. 7 A, left). We observed significantly (P < 0.001) higher CD1a-restricted responses in the presence of PLA2 in a cohort of 18 donors (Fig. 7 A, right). More detailed testing of an individual responder showed that anti-CD1a antibodies prevented the response to CD1a and PLA2 (P < 0.05), confirming the essential role of CD1a (Fig. 7 B, left).


Bee venom processes human skin lipids for presentation by CD1a.

Bourgeois EA, Subramaniam S, Cheng TY, De Jong A, Layre E, Ly D, Salimi M, Legaspi A, Modlin RL, Salio M, Cerundolo V, Moody DB, Ogg G - J. Exp. Med. (2015)

CD1a-restricted reactivity to Apis mellifera venom PLA2 is found among polyclonal T cells from blood of healthy donors. CD3+ cells were isolated by magnetic beads from healthy donor PBMC and cultured for 12–14 d with IL-2 and irradiated K562 cells transfected with CD1a (K562 CD1a) or CD1b (K562 CD1b) or CD1c (K562 CD1c) or CD1d (K562 CD1d) or an empty vector (K562) in the presence of Apis mellifera venom PLA2. CD1 reactivity was then examined by IFN-γ ELISpot with transfected or untransfected K562 cells either in the presence or absence of Apis mellifera venom 100 ng/ml PLA2 (A, left). Data from one representative donor of three are shown. Venom PLA2 specific CD1a-restricted T cell responses were measured in 18 donors (A, right). CD1a-restricted responses were examined in the presence or absence of anti-CD1a (B, left) for donor C1098, and PLA2-neutralizing antibodies (7B, right), and manoalide (specific PLA2 inhibitor) for wasp venom in donor C559 (C, left) and Apis mellifera venom PLA2 in donor C334 (C, right). Representative results are shown from three independent experiments. (D) IFN-γ production was measured by co-incubations of PLA2-specific T cells with mDCs (left) or CD14-derived LC-like DCs (middle) in the presence or absence of anti-CD1a for donor C558. IFN-γ production was measured from co-incubations of PLA2-specific T cells and freshly isolated skin CD1a+ cells in the presence or absence of anti-CD1a or manoalide (right). Representative data from 3 (left and middle) or 2 (right) separate donors are shown. Data are mean of triplicate measurements ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4322046&req=5

fig7: CD1a-restricted reactivity to Apis mellifera venom PLA2 is found among polyclonal T cells from blood of healthy donors. CD3+ cells were isolated by magnetic beads from healthy donor PBMC and cultured for 12–14 d with IL-2 and irradiated K562 cells transfected with CD1a (K562 CD1a) or CD1b (K562 CD1b) or CD1c (K562 CD1c) or CD1d (K562 CD1d) or an empty vector (K562) in the presence of Apis mellifera venom PLA2. CD1 reactivity was then examined by IFN-γ ELISpot with transfected or untransfected K562 cells either in the presence or absence of Apis mellifera venom 100 ng/ml PLA2 (A, left). Data from one representative donor of three are shown. Venom PLA2 specific CD1a-restricted T cell responses were measured in 18 donors (A, right). CD1a-restricted responses were examined in the presence or absence of anti-CD1a (B, left) for donor C1098, and PLA2-neutralizing antibodies (7B, right), and manoalide (specific PLA2 inhibitor) for wasp venom in donor C559 (C, left) and Apis mellifera venom PLA2 in donor C334 (C, right). Representative results are shown from three independent experiments. (D) IFN-γ production was measured by co-incubations of PLA2-specific T cells with mDCs (left) or CD14-derived LC-like DCs (middle) in the presence or absence of anti-CD1a for donor C558. IFN-γ production was measured from co-incubations of PLA2-specific T cells and freshly isolated skin CD1a+ cells in the presence or absence of anti-CD1a or manoalide (right). Representative data from 3 (left and middle) or 2 (right) separate donors are shown. Data are mean of triplicate measurements ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001.
Mentions: Having identified PLA2 as sufficient to generate CD1a-presented antigens in vitro using T cell clones, we next sought to determine if bee venom PLA2 is sufficient to activate polyclonal T cells ex vivo in cellular assays (Fig. 7). As with whole venoms (Fig. 1), we detected higher IFN-γ–producing cells with the addition of 100 ng/ml bee venom PLA2 but only in the presence of CD1a and not other CD1 isoforms (Fig. 7 A, left). We observed significantly (P < 0.001) higher CD1a-restricted responses in the presence of PLA2 in a cohort of 18 donors (Fig. 7 A, right). More detailed testing of an individual responder showed that anti-CD1a antibodies prevented the response to CD1a and PLA2 (P < 0.05), confirming the essential role of CD1a (Fig. 7 B, left).

Bottom Line: Patient studies showed that injected PLA2 generates lysophospholipids within human skin in vivo, and polyclonal T cell responses are dependent on CD1a protein and PLA2.These findings support a previously unknown skin immune response based on T cell recognition of CD1a proteins and lipid neoantigen generated in vivo by phospholipases.The findings have implications for skin barrier sensing by T cells and mechanisms underlying phospholipase-dependent inflammatory skin disease.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Rheumatology, Immunology and Allergy, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, 02114.

Show MeSH
Related in: MedlinePlus