Cryofiltration (CF) was performde for a total of 124 (4-14 months) on six patients with rheumatoid arthritis (RA). In this study, we examined the clinical effectiveness of CF and analyzed removal substances (cryogel) by high performance liquid chromatography (HPLC) and two-dimensional electrophoresis.
A rapid improvement in the arthralgia, morning stiffness and general malaise in all patients was obtained by CF. After CF, one patient could stop steroid therapy and two patients could decrease the maintenance dose of steroid and anti-inflammatory drugs. The therapeutic response to CF was divided into two patterns. One pattern showed a marked rebound phenomenon after each CF at first, then the degree of rebound gradually decreased. The other pattern did not show this marked rebound phenomenon. α
1-globulin, RAHA, IgA, IgM and C3 were significantly different in the rebound phenomenon-negative and -positive groups. However, differences in α
1-globulin and C3 were small, and variance in RAHA was large, and these parameters were not good indicators for distinguishing clinical responses to CF. There was a significant negative correlation between time of plugging the second filter and serum IgM concentration. This means that if the serum IgM concentration becomes high, cryogel in the second filter increases. Therefore, we consider that serum IgM concentration is a good indicator to distinguish these two patterns.
The chromatographic pattern of the cryogel was divided into six peaks, designated A, B, C, D, E and F. Moreover, A peak (high molecular weight substances) was divided into three peaks, A1, A2 and A3. The molecular weights of these three peaks were 1, 300, 000, 1, 000, 000 and 640, 000. Reproducibility was extremely good. We analyzed the same sample by two-dimensional electrophoresis and discovered two abnormal proteins. These substances were very similar to the A and D peaks obtained by HPLC.
We used three kind of columns for HPLC; Protein I-250 and I-125, TSK-G3, 000SW and Asahipak GS-520. Asahipak GS-520 was best for analysis of cryogel. The components of the cryogel did not change during CF therapy and between RA patients, therefore we concluded that the volume of cryogel affects the clinical symptoms of RA.
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