In-vivo impaired T helper 1 cell development in submandibular lymph nodes due to IL-12 deficiency following antigen injection into the anterior chamber of the eye.
OBJECTIVE
To determine the functional properties of antigen-specific T cells that accumulate in lymph nodes following injection of antigen into the anterior chamber (AC).


METHODS
Ovalbumin (OVA)-specific, CD4(+) transgenic T cell receptor (Tcr) T cells (KJ-126(+)) from DO11.10 mice were adoptively transferred into unirradiated syngeneic BALB/c mice who then received an injection of OVA in the AC or subcutaneously (SC). Three days later, KJ126(+) T cells in draining and non-draining lymph nodes were analyzed for clonal expansion, activation, and cell-cycle status by flow cytometry. In addition, the cells' functional phenotype was assessed in vitro by proliferation assays and cytokine production by ELISA.


RESULTS
Tcr T cells localized exclusively to draining lymph nodes after injection of OVA into the AC (neck) and SC (neck, axilla, brachial). In both cases, the KJ126(+) T cells displayed evidence of in-vivo activation and proliferation. T cells from AC-injected donors when stimulated in vitro with OVA produced small amounts of IL-2, but no IFN-gamma, IL-4, IL-5, IL-10, or TGFbeta. By contrast, T cells from SC-injected animals displayed a Th1-like phenotype (IL-2, IFN-gamma). The draining lymph nodes of mice that received systemic or intraocular administration of exogenous IL-12 at the time of the AC injection of OVA also contained KJ126(+) T cells that secreted IFN-gamma and IL-2.


CONCLUSIONS
Antigen attracts antigen-specific T cells to lymph nodes that drain the injection site. Whereas responding T cells in nodes draining SC-injection sites differentiated into Th1-like cells, T cells in nodes draining AC injection failed to differentiated beyond the capacity to secrete IL-2 in response to antigen. Since the failure was reversed by exogenous IL-12, we propose that orderly differentiation of antigen-specific T cells down the Th1 pathway is aborted by the eye, because of a deficit in IL-12 production.
Perez
V L
VL
Department of Ophthalmology, Harvard Medical School, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, Boston, Massachusetts, United States of America.
Biuckians
A J
AJ
Streilein
J W
JW
D019264:Adoptive Transfer; D000818:Animals; D000867:Anterior Chamber / Q000502:physiology*; D000917:Antibody Formation; D002455:Cell Division / Q000187:drug effects; D016922:Cellular Senescence; D007267:Injections; D007371:Interferon-gamma / Q000096:biosynthesis; D018664:Interleukin-12 / Q000172:deficiency* / Q000502:physiology; D008198:Lymph Nodes / Q000166:cytology*; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C; D008822:Mice, Transgenic / Q000235:genetics; D010047:Ovalbumin / Q000008:administration & dosage* / Q000276:immunology / Q000494:pharmacology; D010641:Phenotype; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000378:metabolism; D013363:Submandibular Gland; D006377:T-Lymphocytes, Helper-Inducer / Q000166:cytology / Q000187:drug effects / Q000502:physiology* / Q000637:transplantation
D011948:Receptors, Antigen, T-Cell; D018664:Interleukin-12; D007371:Interferon-gamma; D010047:Ovalbumin