B-Cell-Activating Factor Levels Are Associated With Antibody-Mediated Histological Damage in Kidney Transplantation.

INTRODUCTION: Along with death engraftment, in recent years, antibody-mediated damage has been identified as the leading cause of loss of kidney transplants. Despite the recognition of the role of the B-lymphocyte subpopulation in the development of both tolerance and rejection, little is known about the trigger mechanisms and effectors of this humoral response. BACKGROUND: We analyzed the relationship between B lymphocyte subpopulations and levels of B-cell-activating factor (BAFF) with the histological findings in biopsies of renal transplantation. MATERIAL AND METHODS: We selected 35 patients whose kidney transplant biopsy was performed between January and November 2015. The biopsy specimens were classified according to Banff criteria. At the moment of the biopsy BAFF levels and B-lymphocyte subpopulations in blood were measured using enzyme-linked immunosorbent assay (ELISA) and using flow cytometry, respectively. RESULTS: Mean BAFF levels were 493 ± 245 pg/mL. The median performance of biopsy post-transplantation was 12.9 (11.7-23.9) months. BAFF levels correlated with pretransplantation antibodies (r = 0.523; P = .002) but not with kidney function. In biopsies performed more than 1 year after transplantation BAFF levels correlated with the severity of chronic glomerular (cg) involvement (r = 0.625; P = .003). Histological variables related to antibody-mediated injury selected by principal component analysis (glomerulitis, peritubular capillary, and chronic glomerulopathy) related to BAFF levels (B factor, 116; 95% confidence interval [CI], 12-220; P = .029). Biopsy specimens with transplant glomerulopathy (TG) showed lower levels of circulating naive CD19 + subpopulation, IgD+, and CD27- (32.7 ± 28.1 vs 87.9 ± 79.1; P = .017) compared with biopsy specimens without TG. CONCLUSIONS: Elevated levels of BAFF are associated with increased presence and severity of TG and a set of variables related to antibody-mediated histological damage. TG is associated with changes in circulating B-lymphocyte subpopulations that could contribute to its pathogenesis.

Biological monitoring of isocyanates and related amines. IV. 2,4- and 2,6-toluenediamine in hydrolysed plasma and urine after test-chamber exposure of humans to 2,4- and 2,6-toluene diisocyanate.

Two men were exposed to toluene diisocyanate (TDI) atmospheres at three different air concentrations (ca. 25, 50 and 70 micrograms/m3). The TDI atmospheres were generated by a gas-phase permeation method, and the exposures were performed in an 8-m3 stainless-steel test chamber. The effective exposure period was 4 h. The isomeric composition of the air in the test chamber was 30% 2,4-TDI and 70% 2,6-TDI. The concentration of TDI in air of the test chamber was determined by an HPLC method using the 9-(N-methyl-amino-methyl)-anthracene reagent and by a continuous-monitoring filter-tape instrument. Following the hydrolysis of plasma and urine, the related amines, 2,4-toluenediamine (2,4-TDA) and 2,6-toluenediamine (2,6-TDA), were determined as pentafluoropropionic anhydride (PFPA) derivatives by capillary gas chromatography using selected ion monitoring (SIM) in the electron-impact mode. In plasma, 2,4- and 2,6-TDA showed a rapid-phase elimination half-time of ca. 2-5 h, and that for the slow phase was greater than 6 days. A connection was observed between concentrations of 2,4- and 2,6-TDI in air and the levels of 2,4- and 2,6-TDA in plasma. The cumulated amount of 2,4-TDA excreted in the urine over 24 h was ca. 15%-19% of the estimated inhaled dose of 2,4-TDI, and that of 2,6-TDA was ca. 17%-23% of the inhaled dose of 2,6-TDI. A connection was found between the cumulated (24-h) urinary excretion of 2,4- and 2,6-TDA and the air concentration of 2,4- and 2,6-TDI in the test chamber.(ABSTRACT TRUNCATED AT 250 WORDS)

Biological monitoring of isocyanates and related amines. III. Test chamber exposure of humans to toluene diisocyanate.

Five men were exposed to toluene diisocyanate (TDI) atmospheres for 7.5 h. The TDI atmospheres were generated by a gas-phase permeation method, and the exposures were performed in an 8-m3 stainless-steel test chamber. The mean air concentration of TDI was ca. 40 micrograms/m3, which corresponds to the threshold limit value (TLV) of Sweden. The inhaled doses of 2,4- and 2,6-TDI were ca. 120 micrograms. TDI in the test chamber air was determined by an HPLC method using the 9-(N-methylaminomethyl)-anthracene reagent and by a continuous-monitoring filter-tape instrument. After hydrolysis of plasma and urine, the related amines, 2,4- and 2,6-toluenediamine 2,4-, and 2,6-TDA), were determined as pentafluoropropionic anhydride (PFPA) derivatives by capillary gas-chromatography using selected ion monitoring (SIM) in the electron-impact mode. The urinary elimination of the TDAs showed a possible biphasic pattern, with rapid first phases for 2,4-TDA (mean t1/2 for the concentration in urine, 1.9 h) and for 2,6-TDA (mean t1/2 for the concentration in urine, 1.6 h). The cumulative amount of 2,4-TDA excreted in urine within 28 h ranged from 8% to 14% of the estimated dose of 2,4-TDI, and the cumulative amount of 2,6-TDA in urine ranged from 14% to 18% of the 2,6-TDI dose. The average urinary level of 2,4-TDA was 5 micrograms/l in the 6 to 8-h sample (range 2.8-9.6 micrograms/l), and the corresponding value for 2,6-TDA was 8.6 micrograms/l (range, 5.6-16.6 micrograms/l). Biological monitoring of exposure to 2,4- and 2,6-TDI by analysis of 2,4- and 2,6-TDA in urine is feasible.

Test atmospheres of diisocyanates with special reference to controlled exposure of humans.

An all glass apparatus for the generation of air concentrations of 2,4-toluene diisocyanate (2,4-TDI), 2,6-toluene diisocyanate (2,6-TDI) and 1,6-hexamethylene diisocyanate (HDI) was developed. The generation principle was based on gas-phase permeation with permeation membranes of silicon rubber. In an 8 m3 stainless steel test chamber, low and steady TDI-and HDI atmospheres (1-100 micrograms/m3) could be maintained. The diisocyanate concentrations were determined by an HPLC method, using the 9-(N-methylaminomethyl)-anthracene reagent utilizing UV detection. The sum of diisocyanates and their related amines were determined by sampling in 0.4 M hydrochloric acid solution, and analysis by capillary gas chromatography with thermionic specific detection. Related amines were determined by sampling in ethanol - 0.2% KOH and analysis on GC-TSD. A continuous band-tape monitor was used for the determination of diisocyanates. Losses of diisocyanates in the test chamber were evaluated by measuring the TDI and HDI concentrations at the inlet respectively the outlet of the test chamber. At the outlet of the test chamber, ca 25% of the TDI respectively HDI concentrations were recovered. With a male subject in the test chamber ca 15% of the HDI concentration was recovered. The air flow through the test chamber was ca 10 m3/h. The changes in isomeric composition of airborne TDI, at stopped flow conditions, showed that the decay of the 2,4-isomer was faster than of the 2,6-isomer. No trace of the related amine toluene diamine (TDA) was detected in the test chamber, at TDI concentrations ranging from 20 to 50 micrograms/m3. Sampling losses due to sampling connections were evaluated.(ABSTRACT TRUNCATED AT 250 WORDS)

Trace analysis of airborne aromatic isocyanates and related aminoisocyanates and diamines using high-performance liquid chromatography with ultraviolet and electrochemical detection.

A high-performance liquid chromatographic method was developed for trace analysis of complex air mixtures containing 2,6- and 2,4-toluenediisocyanates and related aminoisocyanates and diamines. The accuracy was tested at isocyanate concentrations of 2-1000 microg/m3 in air. The method is based on derivatization in the sampling step of isocyanate functions to corresponding urethane groups, with alkaline ethanol as the sampling and reacting medium. The derivatives formed, toluenediurethanes and tolueneaminourethanes, and unreacted diamines were detected by UV or electrochemically, the electrochemical detection being one order of magnitude more sensitive. Using an enrichment column, detection limits of ca. 0.05 pg/microl were obtained with electrochemical detection at a potential of 950 mV, which corresponds to air concentrations of 0.1 microg/m3 with 5 min sampling time at a rate of 11/min. The precision in the measurements were ca. 4% at concentrations of 6 microg/m3. A field measurement was performed concerning flame lamination of toluenediisocyanate-based polyurethane and cloth. Isocyanates, aminoisocyanates and diamines were found at air concentrations of 1-100 microg/m3.

Systemic reactions associated with polyisocyanate exposure.

A spray-painter suffered attacks of chills, fever, general malaise, dyspnea and wheezing, headache, arthralgia, and leucocytosis a few hours after exposure to aerosols of varnishes containing two different polyisocyanates based upon monomers of hexamethylene or toluene diisocyanate. Immunologic studies revealed an increase in the serum immunoglobulin G level, but no specific antibodies against isocyanates conjugated to human serum albumin. The polyisocyanate level in the workroom air was high [a time-weighted average of 4.2 mg/m3, corresponding to 17 mumol NCO (isocyanate groups)/m3], the toluene diisocyanate monomer level being much lower (a time-weighted average of 0.03 mg/m3, corresponding to 0.3 mumol NCO/m3).