Antinuclear antibodies and HLA class II alleles in Jamaican patients with systemic lupus erythematosus.

OBJECTIVE: The relationship between human leukocyte antigens class II (HLA) and antinuclear antibodies was investigated in Jamaican patients with Systemic Lupus Erythematosus (SLE). METHODS: Samples of blood of 82 patients with SLE and 75 healthy controls were tested for antinuclear antibodies using the fluorescent antinuclear antibody (FANA) test, counterimmunoelectrophoresis (CIEP) and the Crithidia luciliae immunofluorescence test (CL-IFT). A DNA-based HLA typing method was used to determine the frequencies of alleles of HLA-DRB1, DRB3, DRB4 and DRB5 in patients and healthy controls. RESULTS: The FANA test was positive in all of the sera from patients with SLE. Anti-dsDNA antibodies were present in 49% (40/82), anti-Sm/RNP 44% (36/82) and anti-Ro/La 43% (35/82) of the sera from SLE patients. The frequency of HLA-DR4 was significantly lower in SLE patients than in healthy controls (2/82, 2% vs 15/75, 20%; RR = 0.12; p = 0.0004; CP = 0.005) but no other HLA-DRB1 SLE associations were found. A positive HLA-DR3 anti-Ro/La antibody association was found in the patients with SLE (9/21, 43% vs 5/55, 9%; odds ratio (OR) = 7.5; CP = 0.01). In contrast, possession of HLA-DR6 was negatively associated with the absence of anti-dsDNA antibodies (9/32, 28% vs 27/44, 61%; OR = 0.2; CP = 0.05). CONCLUSION: The HLA-DR6 allele is associated with the absence of antinuclear antibodies and HLA-DR3 with the presence of anti-Ro/La antibodies in Jamaican patients with SLE. However, these results and those of previous studies of Jamaican patients suggest that the HLA-DR3 association with the development of SLE reported in other populations might in fact reflect the association of HLA-DR3 with anti-Ro/La antibodies. Further investigations are needed to determine whether HLA-DRB antinuclear antibody associations define clinical subsets of SLE in Jamaican patients.

Antinuclear antibodies and HLA class II alleles in Jamaican patients with systemic lupus erythematosus / Anticuerpos antinucleares y aleles HLA de clase II en pacientes Jamaicanos con lupus eritematoso sistémico

OBJECTIVE: The relationship between human leukocyte antigens class II (HLA) and antinuclear antibodies was investigated in Jamaican patients with Systemic Lupus Erythematosus (SLE). METHODS: Samples of blood of 82 patients with SLE and 75 healthy controls were tested for antinuclear antibodies using the fluorescent antinuclear antibody (FANA) test, counterimmunoelectrophoresis (CIEP) and the Crithidia luciliae immunofluorescence test (CL-IFT). A DNA-based HLA typing method was used to determine the frequencies of alleles of HLA-DRB1, DRB3, DRB4 and DRB5 in patients and healthy controls. RESULTS: The FANA test was positive in all of the sera from patients with SLE. Anti-dsDNA antibodies were present in 49% (40/82), anti-Sm/RNP 44% (36/82) and anti-Ro/La 43% (35/82) of the sera from SLE patients. The frequency of HLA-DR4 was significantly lower in SLE patients than in healthy controls (2/82, 2% vs 15/75, 20%; RR = 0.12; p = 0.0004; CP = 0.005) but no other HLA-DRB1 SLE associations were found. A positive HLA-DR3 anti-Ro/La antibody association was found in the patients with SLE (9/21, 43% vs 5/55, 9%; odds ratio (OR) = 7.5; CP = 0.01). In contrast, possession of HLA-DR6 was negatively associated with the absence of anti-dsDNA antibodies (9/32, 28% vs 27/44, 61%; OR = 0.2; CP = 0.05). CONCLUSION: The HLA-DR6 allele is associated with the absence of antinuclear antibodies and HLA-DR3 with the presence of anti-Ro/La antibodies in Jamaican patients with SLE. However, these results and those of previous studies of Jamaican patients suggest that the HLA-DR3 association with the development of SLE reported in other populations might in fact reflect the association of HLA-DR3 with anti-Ro/La antibodies. Further investigations are needed to determine whether HLA-DRB antinuclear antibody associations define clinical subsets of SLE in Jamaican patients.

OBJETIVO Se investigó la relación entre los antígenos de leucocito humano (human leukocyte antigens o HLAs). Clase II y los anticuerpos antinucleares en pacientes jamaicanos con lupus eritematoso sistémico (LES). MÉTODOS: Se examinaron muestras de sangre de 82 pacientes con LES y 75 controles saludables para determinar la presencia de anticuerpos antinucleares, usando la prueba del anticuerpo antinuclear fluorescente (FANA), la contrainmunoelectroforesis (CIEP) y el test de inmunofluorescencia con Crithidia luciliae (CL-IFT). Un método de tipificación HLA basado en el ADN fue usado para determinar las frecuencias de aleles de HLA-DRB1, DRB3, DRB4 y DRB5 tanto en los pacientes como en los controles saludables. RESULTADOS: La prueba FANA fue positiva en todos los sueros de pacientes con LES. Anticuerpos anti-dsADN se hallaban presentes en 49% (40/82), anti-Sm/RNP en 44% (36/82) y anti-Ro/La en 43% (35/82) de los sueros de los pacientes de LES. La frecuencia de HLA-DR4 fue significativamente más baja en los pacientes con LES que en los controles saludables (2/82, 2% vs 15/75, 20%; RR = 0.12; p = 0.0004; CP = 0.005) pero no se hallaron otras asociaciones de LES con HLA-DRB1. Se halló una asociación positiva de anticuerpos HLA-DR3 anti-Ro/La en los pacientes con LES (9/21, 43% vs 5/55, 9%; odds ratio (OR) = 7.5; CP = 0.01). En contraste con ello, la posesión de HLA-DR6m estuvo asociada negativamente con la ausencia de anticuerpos anti-dsADN (9/32, 28% vs 27/44, 61%; OR = 0.2; CP = 0.05). CONCLUSIÓN: El alele HLA-DR6 está asociado con la ausencia de anticuerpos antinucleares y el de HLA-DR3 con la presencia de anticuerpos anti-Ro/La en pacientes jamaicanos con LES. Sin embargo, estos resultados al igual que los de los previos estudios de pacientes jamaicanos, sugieren que la asociación HLA-DR3 con el desarrollo de LES reportado en otras poblaciones podría de hecho reflejar la asociación de HLA-DR3 con anticuerpos anti-Ro/La. Se requieren investigaciones ulteriores a fin de determinar si las asociaciones de anticuerpo antinuclear HLA-DRB definen subconjuntos de LES en pacientes jamaicanos.

Counterimmunoelectrophoresis with serum prediffusion: an improved method for the detection and identification of antibodies against extractable nuclear and cytoplasmic antigens.

In the technique of counterimmunoelectrophoresis (CIE) with serum prediffusion (SPD) serum is allowed to diffuse freely into the gel before pouring the antigenic extract in its trough (or wells) and starting the electrophoresis. Both the immunoprecipitations and the interactions with reference sera are strongly intensified by SPD, leading to higher sensitivity and specificity for the detection of anti-SSA/Ro, anti-SSB/La, anti-U1RNP, anti-Sm, anti-Jo1 and even anti-Scl-70 antibodies. We found that the optimal SPD time was 2 h. To evaluate the relevance of SPD for the clinical laboratory, 92 antinuclear antibody (ANA) positive sera were tested on CIE without SPD and with 2 h SPD in identification tests with SSA/Ro, SSB/La, Sm, U1RNP and Jo1 reference sera (rsa). The precipitation lines and their interactions were evaluated by three independent observers. It was observed that SPD considerably improved the efficiency of CIE for antibody identification. The mechanisms underlying the intensification of the precipitation lines by SPD are discussed as are the characteristics of the CIE in comparison with other test systems such as the enzyme linked immunosorbent assay (ELISA) and immunoblot.

Analysis of the stabilizing effect of epsilon-aminocaproic acid by electrophoretic techniques and immunoblotting.

The effect of epsilon-aminocaproic acid (EACA) on the degradation of aqueous pollen extracts was studied by isoelectric focusing, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and immunoblotting. The extracts were stored at 4 degrees C for 7 days and at 37 degrees C for 1, 4, and 7 days. Addition of 0.1 mol/L of EACA before storage partly protected the extract from degradation. The protective effect of EACA could be demonstrated most clearly by immunoblotting, suggesting that more epitopes were preserved in an antigenic configuration. The stabilizing effect increased with higher EACA concentrations.