A Method of Trace Iodination of Proteins for Immunologic Studies

McConahey, Patricia J.; Dixon, Frank J.

doi:10.1159/000229699

Cited by 1,856 publications

(579 citation statements)

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“…The affinity-purified antibodies were labelled with 1 25lodine according to the method of McConahey & Dixon (1966) to a specific activity of 1-5 ,tCi/tg of protein.…”

Section: Methodsmentioning

confidence: 99%

Failure to detect autologous antibodies in the remission sera of patients with AML: complications introduced by the presence of rheumatoid factor

James¹,

Dean²,

Alexander³

1980

Br J Cancer

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Summary.-Sera were collected from patients with acute myelogenous leukaemia (AML) at various times during remission induced by chemotherapy, but after cessation of all immunosuppressive treatment. These sera were tested, by a sensitive assay using radio-labelled antiglobulin binding, for the presence of antibodies which bound to the surface of autologous AML cells. The cell populations examined were chosen on the basis that they proliferated in short-term culture, did not bind anti-Ig reagents directly, and that more than 80% of the cells did not carry detectable Fc receptors. With 8/9 patients studied, no specific antibodies of the IgG or IgM class could be detected in serum samples taken during remission. IgG and IgM antibodies from the remission sera of one patient were found to bind to autologous leukaemic cells, but this was found to be due to the presence of rheumatoid factor (RF) and removal of the RF activity abolished this binding.This study has, like others, failed to detect autologous antibodies, in remission sera, that are directed against membrane components of AML cells.

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“…The affinity-purified antibodies were labelled with 1 25lodine according to the method of McConahey & Dixon (1966) to a specific activity of 1-5 ,tCi/tg of protein.…”

Section: Methodsmentioning

confidence: 99%

Failure to detect autologous antibodies in the remission sera of patients with AML: complications introduced by the presence of rheumatoid factor

James¹,

Dean²,

Alexander³

1980

Br J Cancer

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“…[131I]DNP-BSA was prepared by the reaction of DNP-BSA with Nal31I in the cold by the chloramine T method (27).…”

Section: Methodsmentioning

confidence: 99%

Dual Regulatory Role of the Thymus in the Maturation of Immune Response in the Rabbit

Taniguchi

Tada

1974

The Journal of Experimental Medicine

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Rabbits thymectomized in early adulthood produced more antihapten antibody than sham-thymectomized controls after hyperimmunization with 2,4-dinitrophenyl bovine gamma globulin (DNP-BGG). The average associated constant of anti-DNP antibody produced by thymectomized animals was more than 10 times higher than that of the controls. Similar effects were obtained by extensive treatment of rabbits with antithymocyte serum (ATS) before and during the immunization with DNP-BGG. The results indicated that relative diminution of thymus-derived lymphocytes (T cells) resulted in a stimulation of antibody-forming cells with a higher affinity. On the other hand, preimmunization of rabbits with different doses of BGG caused either enhancement or suppression of the hapten-specific antibody response, depending on the priming dose of BGG. The suppressed antibody response was always associated with a marked decrease in the antibody affinity. If rabbits were partially tolerized with a large dose of soluble BGG, some of the animals produced little antibody against hapten (DNP) coupled to this carrier, and the affinity of produced antibody was low. However, other rabbits tolerized with BGG produced large amounts of anti-DNP antibody upon hyperimmunization with DNP-BGG, whose affinity was only slightly lower than that of the control. These results can be harmonized if it is assumed that the thymus plays an important role in the maturation of the immune response. It is postulated that T cells, in numbers ordinarily available, would first assist in the proliferation of antihapten antibody-forming cell precursors already selected by antigen, thus accounting for the rapid increase of antibody affinity in the early stage of immunization. However, after a larger number of carrier-specific T cells are made in response to continued immunization, these would suppress antibody-forming cells. The suppression would be greater for cells with higher affinity for antigen, resulting in a decrease in antibody affinity. This postulate explains preferential stimulation and suppression of cells having higher affinity receptors under circumstances in which T cell are relatively depleted or overstimulated, and further permits an explanation for the decrease of antibody affinity after long-term immunization.

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“…34,35 Both antibodies were radioiodinated by the chloramine-T method. 36 Protein-bound iodine was separated from free iodine by passage over a PD-10 column (Pharmacia, Piscataway, NJ) equilibrated with 0.04 M phosphate-buffered saline (0.04 M phosphate, 0.15 M NaCl, 0.02% NaN 3 ), pH 7.4, containing 1% human serum albumin. Specific activity of the labeled product was 12-15 Ci/ g immunoglobulin G (IgG).…”

Section: Radioiodinationmentioning

confidence: 99%

Optimizing the use of combined radioimmunotherapy and hypoxic cytotoxin therapy as a function of tumor hypoxia

et al. 2001

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Combined radioimmunotherapy (RAIT) and hypoxic cytotoxin therapy (SR4233 or NLCQ-1) have been evaluated with both modalities administered on the same day with only moderate improvement compared with the effects of RAIT alone. In a series of studies using oxygen electrodes, immunohistochemistry and radiotracers, we have demonstrated that RAIT induces a prolonged state of hypoxia in most tumors, without affecting the pO 2 levels in normal tissues. Using serial microelectrode measurements through subcutaneous (s.c.) GW-39 human colonic xenografts, we established that the median pO 2 was unrelated to the initial size of the tumor, over a range of sizes from 1.0 to 4.0 cm. Fourteen days after mice were given a 240-Ci dose of 131 I-MN-14 anti-carcinoembryonic antigen immunoglobulin G, their median pO 2 declined from 26.1 ؎ 9.6 mmHg to 9.8 ؎ 3.9 mmHg (p < 0.001). Using the radiotracer 3 H-MISO that accumulates in hypoxic regions, uptake in GW-39, LoVo and LS174T s.c. human colonic tumors increased 3.0-to 4.2-fold from day 14 through day 28 post-RAIT, but uptake of 3 H-MISO in CALU-3 tumors remained unchanged after RAIT. Normal tissue (liver, kidney, lung) uptake of 3 H-MISO did not exhibit significant changes. The increase in tumor hypoxia was also demonstrated visually using anti-PIMO staining of tumor sections. We postulated that sequential delivery of the 2 therapeutic agents, with the hypoxic cytotoxin given 2 weeks after RAIT when tumor pO 2 levels were at their nadir, would improve the therapeutic response above either modality alone or above the 2 agents delivered on the same day. Tumor growth was compared in mice given either RAIT or cytotoxin alone, the combined treatment on the same day or with the cytotoxin delivered 14 days after RAIT. Tumor size on day 35 for RAIT-treated and SR4233-treated GW-39 were 3.56 ؎ 0.40 and 7.98 ؎ 2.50 cm 3 . When RAIT ؉ SR4233 were delivered on the same day, tumor size dropped to 2.78 ؎ 0.80 cm 3 . If RAIT was given on day 0 and SR4233 on day 14, size further declined further to 1.74 ؎ 0.32 cm 3 (p < 0.05 compared with same day delivery). For LS174T, tumor size on day 28 for RAIT-treated and SR4233-treated tumors were 1.14 ؎ 0.36 cm 3 and 3.65 ؎ 0.78 cm 3 , respectively. When RAIT ؉ SR4233 were delivered on the same day, size was 0.51 ؎ 0.174 cm 3 . If RAIT was dosed on day 0 and SR4233 was given on day 14, tumor size was 0.13 ؎ 0.07 cm 3 (p < 0.05). Similar results were obtained for LoVo, but not for CALU-3 tumors. Another hypoxic cytotoxin, NLCQ-1, was also more efficacious 2 weeks after RAIT, compared with same-day dosing. Thus, information on tumor hypoxia after radioantibody therapy could be important for ascertaining a window of opportunity when the surviving tumor regions are most responsive to hypoxic cytotoxins.

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A Method of Trace Iodination of Proteins for Immunologic Studies

Cited by 1,856 publications

References 1 publication

Failure to detect autologous antibodies in the remission sera of patients with AML: complications introduced by the presence of rheumatoid factor

Failure to detect autologous antibodies in the remission sera of patients with AML: complications introduced by the presence of rheumatoid factor

Dual Regulatory Role of the Thymus in the Maturation of Immune Response in the Rabbit

Optimizing the use of combined radioimmunotherapy and hypoxic cytotoxin therapy as a function of tumor hypoxia

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