A Radioisotopic Method to Measure Delayed Type Hypersensitivity in the Mouse

Abstract

Sensitized lymphoid cells could transfer to normal non-sensitized (naive) mice by 24-48 h after antigen challenge in the ear, the capacity to incorporate, at the site of antigen deposition, 5-iodo-2′deoxyuridine-¹²⁵I in amounts significantly above those obtained in control mice. This was associated with a mononuclear cell infiltration in the pinna. In contrast to lymphoid cells, serum antibodies were unable to transfer a 24- or 48 hour ear reaction. The cells responsible were T lymphocytes as demonstrated by successful transfer following enrichment for T lymphocytes, and abrogation of transfer following treatment with anti-Θ serum and complement. Transfer was achieved whether the naive recipients were normal, T-cell deprived, pretreated with cyclophosphamide, or lightly irradiated but not when they were heavily irradiated. Adoptive transfer of the 24-hour ear response was demonstrated with three different antigenic systems. The time-response curves were different with each system although peak reactions were obtained 5 days after sensitization of the donors in all cases. The specificity patterns of the 24-hour ear reactions on transfer were similar to those obtained in the sensitized donors. The results of these studies indicate that the radioisotopic ear method can, under defined conditions, demonstrate the existence of a state of delayed type hypersensitivity in the donors.

H-2 gene complex restricts transfer of delayed-type hypersensitivity in mice.

“…The test used for DTH has been described in detail elsewhere (11,12). It depends on the labeling of replicating cells by '25I-dUrd and the quantification of the influx of labeled mononuclear cells (presumably monocytes) into a defined site where antigen is deposited as a challenge (left ear).…”

“…The present investigations were designed to determine whether successful transfer of DTH by lymphoid cells from sensitized to naive mice might be restricted by genes of the MHC. These studies were made possible by the use of congenic strains of mice and of a new radioisotopic assay which gives an objective measure of the extent of DTH in mice (11,12).MATERIALS AND METHODS Mice. Female mice (2-3 month old) were used for sensitization.…”

“…Mice of both sexes served as recipients of sensitized cells. Highly inbred specific pathogen-free CBA/CaHWehi, C57BL/6JWehi, and BALB/cAnBrWehi mice were transferred to conventional rooms for the duration of the experi- (2) it is associated predominantly with a mononuclear cell infiltration; (3) it is not evident in T-cell-deprived mice; (4) it is not given by antigens unable to activate T cells, and (5) the capacity to elicit the 24-or 48-hr ear response can be transferred from sensitized to naive mice by purified T lymphocytes, not by serum antibodies (11,12). Values are arithmetic means + 1 SEM.…”

Role of major histocompatibility complex gene products in delayed-type hypersensitivity.

“…Specific lysis of virusinfected or chemically modified target cells occurred effectively only when the cytotoxic T lymphocytes and their targets were MHC compatible, the genes required for this interaction mapping in the K or D, but not I, regions (4,5). By contrast, transfer of delayed-type hypersensitivity (DTH) to protein antigens by sensitized T cells was successful only in I-A compatible recipients (6). The inability to transfer DTH in incompatible recipients could not be attributed to rejection of the injected cells, to their total recruitment into areas such as the spleen, nor to their engagement in a mixed lymphocyte reaction (6).…”

“…By contrast, transfer of delayed-type hypersensitivity (DTH) to protein antigens by sensitized T cells was successful only in I-A compatible recipients (6). The inability to transfer DTH in incompatible recipients could not be attributed to rejection of the injected cells, to their total recruitment into areas such as the spleen, nor to their engagement in a mixed lymphocyte reaction (6). Thus, for example, DTH was successfully transferred from Abbreviations: T lymphocytes, thymus-derived lymphocytes; B cell, bone-marrow-derived lymphocyte; DNFB, 2,4-dinitro-1-fluorobenzene; DTH, delayed-type hypersensitivity; FGG, fowl gamma globulin; 125I-dUrd, 5-[12Iliodo-2'-deoxyuridine; L/R l25I-dUrd uptake, ratio of radioactivity of left ear to radioactivity of right ear; MHC, major histocompatibility complex; TBMC, tetraparental bone marrow chimera.…”