Comparison of Murine Delayed Hypersensitivity Reactions Elicited with Particle-Associated Versus Soluble Human γ-Globulin

Abstract

Footpad reactions elicited in DHS mice using human γ-globulin (HGG)-coated polystyrene latex particles and soluble HGG (sHGG) were compared. Both the magnitude and the persistence of DHS lesions produced by HGG-latex were considerably greater; at 24, 48 and 72 h after challenge, the level of reactivity induced by HGG-latex was 24, 41 and 71% higher, respectively, than those elicited with sHGG. Early nonspecific swelling following injection of HGG-latex in footpads of normal mice was negligible by 24 h, whereas Arthus-responsive animals did not return to control levels until 48 h. The possible advantages and limitations of using particle-associated protein to elicit DHS reactions are discussed.

A Method for Eliciting Indurated DTH Reactions to Soluble Protein Antigens in the Flank Skin of Mice: Correlation of Visual Measurements with ¹²⁵I-UdR Uptake Indices

“…A l though the immunizing dose, route of im munization and the method used to assess D T H in our studies differed from those of other investigators [3,9,10], our results are in general agreement with their findings. Treatment of mice with cyclophosphamide prior to immunization has been reported by Vadas et al [12] to result in early (day 5) and enhanced expression of D T H .…”

“…Relatively small immunizing doses of an tigen (10/(g) were used in the present study, while other investigators have used 50 «g [10], 500 ,ug [12], and 1.000 »g [3,6,9] of soluble protein antigens in C F A for immu nization. When large (1,000/rg) doses of antigen were used for immunization, strong 3-to 4-hour Arthus type reactions in the footpads were reported by Katsura et al [6], There was a sharp decline in these early reactions by 8 h after antigen challenge, and this was followed by the development of D TH reactions 24-48 h later in the same site.…”

Characterization and Quantitation of Murine-Delayed Hypersensitivity Responses Elicited with Particle-Associated Human γ-Globulin

“…On day 11, however, data obtained in 4 separate experiments in dicated that CY + ID treated mice retained a mean of 73°/o of the 10 tig ID dose which was shown to be retained by control ani mals; thus, only marginal antibody produc tion in the sensitized groups had occurred (data not shown). When the effect of varying the challenge dose of ,25I-HGG-La on the magnitude of the DHS reactions produced was examined, the data (table II) Although our previous report of DHS responses elicited with HGG-La, as opposed to sHGG, indicated that the lesions were more pronounced and of greater duration, the reason for this phenomenon was not ex plored [3]. The possibility that stronger DHS reactions may be induced by particu late antigens because of increased retention time at the site of challenge was examined using radiolabeled, particle-associated and soluble HGG.…”

“…The question of whether the molecular form of an antigen has an impact on the magnitude of the delayed response was prompted by the suggestion that particulate antigens such as erythrocytes [14], tumor cells [8], viruses, flagellin, and hcmocyanin [61 produce more marked DHS lesions than do noncationic soluble molecules [7], In an earlier study we found that particle-asso ciated rather than soluble HGG was in fact much more effective in eliciting murine DHS reactions; the lesions were of greater magnitude and more persistent [3], The cur rent study extends these observations and characterizes the 'HGG system' with respect to the onset of antibody production, the his tologic components involved in the lesions produced, the conditions for HGG coating of latex particles, the dose-response rela tionship following challenge with HGG-La, and the persistence of particle-associated as opposed to soluble HGG at the site of chal lenge.…”

“…In a previous study [3], we found that SJL/J mice sensitized with aggregated hu man y-globulin (AHGG) in complete Freund's adjuvant (CFA) developed more marked and persistent delayed hypersensi tivity (DHS) reactions if they were chal lenged with latex particle-associated HGG (HGG-La) rather than soluble HGG (sHGG). This comparison was prompted by the suggestion that particulate antigens [6] provoked much stronger DHS lesions than soluble protein molecules [7j.…”