D8/17 and CD19 expression on lymphocytes of patients with acute rheumatic fever and Tourette's disorder

Abstract

D8/17, an alloantigen found on B lymphocytes, has been reported to be elevated in patients susceptible to rheumatic fever and may be associated with autoimmune types of neuropsychiatric disorders. The pediatric-autoimmune-neuropsychiatric-disorders-associated-with-streptococci model is a putative model of pathogenesis for a group of children whose symptoms of obsessive-compulsive disorder and Tourette's disorder (TD) are abrupt and may be triggered by an infection with group A streptococci. As a test of this model, we have examined D8/17 levels on the B cells of patients with TD and acute rheumatic fever (ARF) along with those on the B cells of normal controls by flow cytometry. We have utilized several different preparations of D8/17 antibody along with a variety of secondary antibodies but have been unable to show an association with an elevated percentage of D8/17-positive, CD19-positive B cells in either ARF or TD. We did find, however, that the percentages of CD19-positive B cells in ARF and TD patients were significantly elevated compared to those in normal controls. Group A streptococcal pharyngitis patients also had an elevated percentage of CD19 B cells, however. These studies failed to confirm the utility of determining the percentage of B cells expressing the D8/17 alloantigen in ARF patients or our sample of TD patients. In contrast, the percentage of CD19-positive B cells was significantly elevated in ARF and TD patients, as well as group A streptococcal pharyngitis patients, suggesting a role for inflammation and/or autoimmunity in the pathogenesis of these disorders.

FIG. 1.

Percentage of total lymphocytes expressing D8/17 versus percentage of CD19-positive B cells expressing D8/17. Flow cytometry histograms for a normal control, a TD patient, and an ARF patient show the different results obtained by analyzing the percentage of total lymphocytes expressing D8/17 (A) and the percentage of CD19-positive B cells expressing D8/17 (B). D8/17 antibody was from Goodwin (7.5 μg/ml). PE, phycoerythrin.

FIG. 2.

Percentage of cells in positive (VWB) cell line versus percentage of cells in negative (LG2) cell line expressing D8/17 antigen. D8/17 antibody was from the ATCC (3 μg/ml). Bar graphs compare the mean percentages of D8/17-positive B cells in the VWB cell line (mean ± SD, 19.79% ± 10.93%; n = 19) and the LG2 cell line (mean ± SD, 2.09% ± 1.24%; n = 19; P = 0.000001). SE, standard error.

FIG. 3.

Flow cytometry histograms comparing mouse IgM negative control (3 μg/ml; Caltag) and D8/17 monoclonal antibody (3 μg/ml; ATCC) expression on CD19-positive B cells from the same subject. PE, phycoerythrin.

FIG. 4.

Percentage of total lymphocytes expressing the CD19 B cell marker in ARF, TD, and pharyngitis patients and controls. The dot plot shows the differences among 89 controls (mean ± SD, 11.106% ± 2.52%), 23 ARF patients (mean ± SD, 18.01% ± 3.99%; P, <0.00000002 against normal controls), 33 TD patients (mean ± SD, 17.70% ± 5.95%; P, 0.0000005 against normal controls and 0.819 against ARF patients), and 17 pharyngitis patients (mean ± SD, 15.25% ± 4.38%; P, <0.002 against normal controls, 0.054 against ARF patients, and 0.115 against TD patients).

FIG. 5.

ASO and anti-DNase B titers in ARF, TD, and pharyngitis patients. The dot plot shows the differences in titers among 17 ARF patients (mean ASO titer, 412 ± 119 IU/ml; mean anti-DNase B titer, 1,002 ± 228), 27 TD patients (mean ASO titer, 163 ± 50 IU/ml [P, <0.07 against ARF patients]; mean anti-DNase B titer, 267 ± 70 [P, 0.006 against ARF patients]), and 17 pharyngitis patients (mean ASO titer, 153 ± 40 IU/ml [P, 0.054 against ARF patients and 0.877 against TD patients]; mean anti-DNase B titer, 250 ± 59 [P, 0.005 against ARF patients and 0.859 against TD patients]).