Measurement of CD4+ and CD8+ T-Lymphocyte Cytokine Secretion and Gene Expression Changes in p-Phenylenediamine Allergic Patients and Tolerant Individuals

Abstract

Factors predisposing to individual susceptibility to contact allergic dermatitis are ill defined. This study was designed to characterize the response of allergic and tolerant individuals' T-lymphocytes after exposure to p-phenylenediamine (PPD). Peripheral blood mononuclear cells (PBMCs) from allergic patients proliferated when treated with PPD and Bandrowski's base (BB) and secreted IL-1alpha, -1beta, -4, -5, -6, -8, -10, and -13; IFN-gamma; tumor necrosis factor-alpha; MIP-1alpha/beta; MCP-1 (monocyte chemotactic protein-1); and RANTES. PBMCs from tolerant individuals were stimulated to proliferate only with BB, and they secreted significantly lower levels of Th2 cytokines. Principal component analysis showed that genes are differentially expressed between the patient groups. A network-based analysis of microarray data showed upregulation of T helper type 2 (Th2) gene pathways, including IL-9, in allergic patients, but a regulatory gene profile in tolerant individuals. Real-time PCR confirmed the observed increase in Th2 cytokine gene transcription in allergic patients. Purified CD4+ and CD8+ T cells from allergic patients were stimulated to proliferate and secrete Th2 cytokines following antigen exposure. Only CD4+ T cells from tolerant individuals were stimulated by BB, and levels of Th2 cytokines were 80% lower. The nature of the antigenic determinant stimulating PBMCs and levels of Th2 cytokines, including IL-9, was confirmed in a validation cohort. These studies show increased activity of Th2 cytokines in CD4+ and CD8+ T cells from individuals with allergic contact dermatitis.

Idiosyncratic Adverse Drug Reactions: Current Concepts

“…PPD-specific T-cell responses were not detected. Bandrowski's base-responsive T cells are also detected in hair dye-exposed human subjects; however, their presence seems to reflect an acquired immune response that does not translate into an allergic reaction (Sieben et al, 2002;Coulter et al, 2008Coulter et al, , 2010. A second population of Th2 secreting T cells, which are stimulated with PPD-derived primary oxidation products but not Bandrowski's base, are detected in patients with hair dye allergy, and their presence seems to represent an important discriminator between allergic and tolerant patient groups.…”

Severe facial swelling in a pregnant woman after using hair dye

“…In addition, the number of pregnant women who dye their hair will also increase, with potential risks for the mother and child. Individuals with allergic contact dermatitis after exposure to PPD demonstrate increased activity of Th2 cytokines in CD4 and CD8 T cells 8. A maternal non-respiratory allergic response during pregnancy, especially where there is increased activity of Th2 cytokines such as occurs in PPD allergy, can mediate the maternal transfer of asthma risk to children 9.…”

The lymphocyte transformation test in allergic contact dermatitis: New opportunities

“…Measurement of induced secretion of IFNg and IL-12 provide markers of selective Type 1 (T H 1/Tc1) responses, whereas IL-4, -5, -10 and -13 can be used as markers of preferential Type 2 (T H 2/Tc2) responses. Subjects allergic to the contact allergen PPD displayed increased activity of Type 2 cytokines in both CD4 and CD8 T-lymphocytes in LTT assays (Coulter et al, 2010). In response to a mixture of MCI and MI, cellular proliferation has been characterized by secretion of both T H 1 (IL-2 and IFNg) and T H 2 (IL-4, -5 and -13) cytokines .…”

“…As an alternative to unfractionated PBMC, purified CD8 and CD4 T-lymphocyte populations have been used in the LTT and cultured with autologous monocyte-derived DC to assess the cytokine secretion profiles of each subset in PPD allergic patients (Coulter et al, 2010). Although the use of purified T-lymphocyte subsets has the advantage of allowing assessment of the responsiveness of a particular cell subset, it is does not provide an holistic reflection of the immune response directed against a chemical allergen.…”

“…Following this early use in the study of nickel sensitized patients, the LTT was further developed to characterize allergenspecific responses to other contact allergens, including metal salts such as cobalt , chromium (Rasanen et al, 1991), MCI (methylchloroisothiazolinone) , DNCB (2,4-dinitrochlorobenzene) (Pickard et al, 2007), and PPD (Coulter et al, 2010;Kimber et al, 1990;Kneilling et al, 2010;McFadden et al, 2011). Although metals such as nickel can, for the purposes of the LTT, be readily dissolved in aqueous culture media in the form of metal salts, difficulties arise when using lipophilic chemicals (which applies to most contact allergens).…”