Affinity columns containing anti-DNA Id+ human myeloma proteins adsorb human epibodies from intravenous gamma globulin.
Affinity columns containing anti-DNA Id+ human myeloma proteins adsorb human epibodies from intravenous gamma globulin.
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含有抗 DNA Id 人骨髓瘤蛋白的亲和柱从静脉内丙种球蛋白中吸附人表观体。
DOI：
10.1002/art.1780400413
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发表时间：
1997
影响因子：
--
通讯作者：
Silvestris,F
中科院分区：
文献类型：
--
作者：
WilliamsJr,RC;Malone,CC;Fry,G;Silvestris,F
目的。研究由带有抗 DNA 独特型 (Id) 标记 16/6、F4、3I 和 8.12 的人阳离子 IgG 骨髓瘤蛋白亲和柱制备的静脉注射丙种球蛋白 (IVGG) 的洗脱液，以了解可能的抗 Id（结合位点）阻断活性。方法。在 3 种含有 IVGG 的制剂中研究了抗 DNA 独特型抗体活性。 高、中、低水平的 IgG 抗 F(ab')2，以及 4 种其他商业 IVGG 制剂。来自系统性红斑狼疮 (SLE) 患者的亲和纯化 IgG 抗 DNA (APAD) 被生物素化，并与酶联免疫吸附测定板上包被的 DNA 结合，用于测量抗 DNA 抗体活性。 IVGG 被吸附到与一组抗 DNA Id 标记物 16/6、F4、3I 和 8.12 呈阳性的阳离子人 IgG 骨髓瘤蛋白连接的 Sepharose 4B 亲和柱上。吸附到此类柱上的材料在低 pH (2.5) 下洗脱，中和后测试其抑制生物素化 APAD 与 DNA 反应的能力。结果。只有 0.05–0.9% 的 IVGG 牢固地结合到 Id 亲和柱上。然后使用 pH 2.5 甘氨酸盐水洗脱这些 IVGG，并将洗脱液中和至 pH 7.4。比较柱流出液和洗脱液组分阻止 SLE APAD 与 DNA 反应的能力。用抗 DNA Id 亲和柱的洗脱液观察到 SLE APAD 结合位点的显着抑制；然而，IVGG 抗 F(ab')2 活性与 APAD 的真正抗 Id 阻断之间没有明显相关性。柱流出物中没有残留的抗Id活性。对于来自不含 4 种抗 DNA Id 标记的人阳离子骨髓瘤柱的 IVGG 洗脱液，没有记录到抗 Id 阻断活性。 IVGG 或 Id 柱洗脱液的 DNase 处理不影响抗 Id 阻断活性。因此，所有可检测的抗 DNA 独特型抗体都能够阻断与 Id+ 亲和柱结合的 SLE 抗 DNA 结合位点。柱洗脱液还显示出一定相对浓度的 IgG 抗 DNA 活性，其对 DNA 的亲和力低于洗脱液中存在的阻断抗 DNA 结合位点的抗体。结论。人抗 DNA Id 柱洗脱液中同时存在抗 DNA 和抗独特型（抗结合位点）活性，表明使用此策略时，来自 IVGG 的表观体相对集中。这种方法可能会带来治疗 SLE 肾炎的新策略。
Objective.To study eluates of intravenous gamma globulin (IVGG) prepared from affinity columns of human cationic IgG myeloma proteins bearing anti‐DNA idiotype (Id) markers 16/6, F4, 3I, and 8.12 for possible anti‐Id (combining site) blocking activity.Methods.Anti‐DNA idiotypic antibody activity was studied in 3 preparations of IVGG containing high, medium, and low levels of IgG anti‐F(ab′)2, and in 4 other commercial IVGG preparations. Affinity‐purified IgG anti‐DNA (APAD) from systemic lupus erythematosus (SLE) patients was biotinylated, and binding to DNA coated on enzyme‐linked immunosorbent assay plates was used to measure anti‐DNA antibody activity. IVGG was adsorbed to Sepharose 4B affinity columns linked to a panel of cationic human IgG myeloma proteins positive for anti‐DNA Id markers 16/6, F4, 3I, and 8.12. Material adsorbing to such columns was eluted at low pH (2.5) and after neutralization, tested for its ability to inhibit biotinylated APAD reacting with DNA.Results.Only 0.05–0.9% of IVGGs bound firmly to Id affinity columns. These IVGGs were then eluted, using pH 2.5 glycine‐saline and eluates neutralized to pH 7.4. Column flowthrough and eluate fractions were compared for their ability to block SLE APAD reacting with DNA. Significant inhibition of SLE APAD combining sites was observed with eluates from anti‐DNA Id affinity columns; however, no correlation between IVGG anti‐F(ab′)2activity and true anti‐Id blocking of APAD was apparent. No residual anti‐Id activity remained in column flowthrough fractions. No anti‐Id blocking activity was recorded for IVGG eluates from human cationic myeloma columns devoid of the 4 anti‐DNA Id markers. DNase treatment of IVGG or Id column eluates did not affect anti‐Id blocking activity. Thus, all detectable anti‐DNA idiotypic antibody capable of blocking SLE anti‐DNA combining sites bound to Id+ affinity columns. Column eluates also showed some relative concentration of IgG anti‐DNA activity, which was of lower affinity for DNA than antibodies also present in eluates which blocked anti‐DNA combining sites.Conclusion.The presence of both anti‐DNA and antiidiotypic (anti‐combining site) activity in human anti‐DNA Id column eluates indicates that epibodies from IVGG are relatively concentrated when this strategy is used. This approach may lead to a new strategy for treatment of SLE nephritis.
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DOI：
--
发表时间：
1984
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Halpern,R;Schiffenbauer,J;Solomon,G;Diamond,B
通讯作者：
Diamond,B
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DOI：
10.1016/0090-1229(91)90088-r
发表时间：
1991
期刊：
Clinical immunology and immunopathology
影响因子：
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作者：
Silvestris,F;Rots,N;YanceyJr,WB;Malone,C;Searles,R;Solomon,A;Dammacco,F;WilliamsJr,RC
通讯作者：
WilliamsJr,RC
人类免疫缺陷病毒-1感染个体中自身抗体识别的独特型肽的鉴定。
DOI：
10.1172/jci118122
发表时间：
1995
期刊：
The Journal of clinical investigation.
影响因子：
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作者：
Wang,QL;Wang,HT;Blalock,E;Muller,S;Kohler,H
通讯作者：
Kohler,H
影响因子：
--
作者：
F. Silvestris;A. Bankhurst;R. Searles;R. Williams
通讯作者：
F. Silvestris;A. Bankhurst;R. Searles;R. Williams
F4、8.12、3I 和 16/6 抗 DNA 独特型相关抗原在阳离子人 IgG 骨髓瘤蛋白上的表达。
DOI：
10.1006/clin.1994.1190
发表时间：
1994
期刊：
Clinical immunology and immunopathology
影响因子：
--
作者：
R. Williams;A. Schriber;C. Malone;F. Silvestris;N. Hannigan;M. Klein;C. Namyst;R. Kyle
通讯作者：
R. Kyle