In Vitro Lymphocyte Responses in Contact Hypersensitivity

John E. Milner
1970 Journal of Investigative Dermatology  
Contact hypersensitivity to dini t rofluorobenzene (DNFB) was developed il1 gu inea pigs. Lymphocytes from sensit ive or insensit ive dono rs were obtain ed from lymph nodes and cultured in the presence of conjugates of DNFB and p rotein extracts of guineCl. p ig epid ermis. Transformation of lymphocytes was assessed by uptake of t rit iated t hyrn idin c by nu cleic acids, and indi cated that lymphocytes from sensit ized dono rs responded in vitro to a greater deg ree t han t hose from
more » ... ized donors. At t he present time, the diagnosis of contact dermaLiLis is established by a 19 th century tec hnique, t he patch test (1). Alt hough considerably refined, the test is essentially the same as it was 70 years ago and retains the sn me hazards such as exacerbation of a spreading dermatitis and inadvertent sensitization of t he patient. As t he only available immunologic test of contact dermatitis, it is still widely used for this pu rpose. Since contact dermatitis is mediated by cells (lymphocytes) rather than circulating antibodies, study of this phenomenon in vitro has been difficul t . In contradistinction, circulating ant ibodies have been studied e},'i;ensively in vitro with quantitation in terms of hemagglutin a nd precipitin titers, complement fixation, etc. None of t hese techniques has been applicable to quant itation of t he immune capacity of the lymphocyte. In 1960, Nowell reported that lymphocytes in tissue culture enlarge (transform) into blast cells and divide in the presence of phytohemagglut inin (2). Although phytohemagglutinin and pokeweed mitogen (3) are non-specific stimulators of lymphocytes, other workers fo und simil ar responses to a number of antigens such as t uberculin , smallpox vaccine, oidomycin, etc ., ( 4, 5, 6 ). In t heir hands t his technique This slud~· supported by Burenu of Occupational Safety and H ealt h (Environmental Control Administration) Grrmt. No. R 01 EC 00238-02. Received January 9, 1970; accepted fo r publicatio n F ebrua ry 20, 1970. *From Lhe Departments of Preventive M edicin e (Envi ronmental Health Division) an d Medicin e (Dermatology). University of \¥ashington , SeaLUe. Washington 98105. ' I' NaLional Cancer Instilute Resea rch Caree r D evelopment Award ee. l-K4 CA 33563-01 , IMB. has proven to b e a specific and reliable measure of a donor's sensitivity to t he antigen tested. Recent work indicates t hat this antigen-indu ced lym phocyte transformation is st rongly correlated with delayed hypersensitivity (7) , and t hus gives promise of developing into a method of in vitro quantitation of the immune capacity of lymphocytes mediating delayed hypersensitivity reactions. Since contact dermatitis appears to be the result of delayed hypersensitivity toward a hapten-skin protein conjugate (8, 9), lymphocytes from sensitized donors should transform in t he presence of appropriate conjugates. If this proved to be the case, lymphocyte t ransformation should not only provide additional information about t he basic mechanisms of contact hypersensitivity, but could also serve as an in vitro method for diagnosing contact dermat itis which would preclude t he wellknown hazards of patch testing. The intent of t his study, therefore, was to determine if lymphocytes from guinea pigs wit h contact hypersensitivity to dinitrofluorobenzene (DNFB) would react in vitro to conjugates of DNFB and skin proteins. Sin ce uptake of t ritiated t hymidine by nucleic acids of lymphocytes in culture has been found to correlat e well with t ransformation into blast cells (10) , t his method was used to evaluate the reactions. 34. MATE RIALS AND METHODS I mmuniza tions. Randomly bred m ale guinea pigs weighing 400 to 600 grams were used . They were individually caged and rn aintained on P urina C how with da ily leLtuce supplementa tion.
doi:10.1111/1523-1747.ep12290518 pmid:5425064 fatcat:xwxykou4nzd27peljm3aqleq2e