title: Study of T-cell activation in type I diabetic patients and pre-type I diabetic subjects by cytometric analysis: antigen expression defect in vitro.
abstract: In Type I diabetes the observation of a decreased release of interleukin-2 (IL-2) and soluble IL-2 receptors by means of stimulated lymphocytes in vitro indicates that a primary immunoregulatory defect may be involved. To confirm this hypothesis we investigated the T-cell activation trend, evaluating the surface expression of IL-2 receptor (CD25), transferrin (CD71), HLA class II (DR), and CD69 phenotypes after in vitro stimulation with phytohemagglutinin (PHA; 1 and 10 micrograms/ml) and concanavalin A (12.5 micrograms/ml) in six newly diagnosed Type I diabetics and six islet cell- and insulin autoantibody-positive first-degree relatives. As controls were studied six long-standing Type I diabetics and six healthy subjects. T-cell cultures from the four groups were performed on the same day and examined at 0, 24, 48, 96, 120, and 144 hr. Cytometric analysis was performed, keeping PBMC gating constant on the basis of physical parameters (scatter and volume). Using both PHA concentrations, a lower level of CD25, CD71, CD69, and DR antigen expression was found in newly diagnosed patients at all observation times with respect to control cultures (P < 0.001). Unexpectedly, pre-Type I diabetic subjects, after 1 microgram/ml of PHA, showed a significantly reduced expression of CD69 (P < 0.001) and CD71 (P < 0.001). The levels remained low, also with high PHA, at the different observation periods, while CD25 expression was found to be reduced in prediabetics only after 1 micrograms/ml of PHA (P < 0.001). The long-standing patients showed a T cell activation trend very close to the latter. Our data show that in Type I diabetes and in the early phases of the disease, the initial activation signal(s) appears to be affected, particularly with one or more subsequent events necessary to initiate the appearance of "activation antigens." This study suggests that the natural history of immunoregulation in pre-Type I and Type I diabetes is characterized by a primary defect in this system, which also persists in patients with long-standing disease.
authors.affiliation: Laboratory of Immunology, University of Palermo, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Giordano
authors.forename: R
authors.initials: R
authors.lastname: De Maria
authors.forename: M
authors.initials: M
authors.lastname: Todaro
authors.forename: G
authors.initials: G
authors.lastname: Stassi
authors.forename: A
authors.initials: A
authors.lastname: Mattina
authors.forename: P
authors.initials: P
authors.lastname: Richiusa
authors.forename: G
authors.initials: G
authors.lastname: Galluzzo
authors.forename: F
authors.initials: F
authors.lastname: Pantó
authors.forename: A
authors.initials: A
authors.lastname: Galluzzo
mesh_terms: D000293:Adolescent; D000328:Adult; D015703:Antigens, CD / Q000032:analysis / Q000502:physiology; D000944:Antigens, Differentiation, B-Lymphocyte / Q000032:analysis / Q000502:physiology; D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis / Q000502:physiology; D017252:CD3 Complex / Q000032:analysis; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D016827:CD8 Antigens / Q000032:analysis; D003922:Diabetes Mellitus, Type 1 / Q000276:immunology*; D005260:Female; D006684:HLA-DR Antigens / Q000032:analysis; D006801:Humans; D007376:Interleukin-2 / Q000378:metabolism; D007700:Kinetics; D037181:Lectins, C-Type; D008213:Lymphocyte Activation; D008297:Male; D010835:Phytohemagglutinins / Q000494:pharmacology; D015375:Receptors, Interleukin-2 / Q000032:analysis / Q000502:physiology; D011990:Receptors, Transferrin; D013601:T-Lymphocytes / Q000276:immunology*; D013997:Time Factors; D000099192:CD69 Antigens
chemical_list: D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; D000945:Antigens, Differentiation, T-Lymphocyte; D017252:CD3 Complex; D016827:CD8 Antigens; D006684:HLA-DR Antigens; D007376:Interleukin-2; D037181:Lectins, C-Type; D010835:Phytohemagglutinins; D015375:Receptors, Interleukin-2; D011990:Receptors, Transferrin; D000099192:CD69 Antigens; C068126:CD71 antigen