In vivo effects in mice of an anti-T cell immunotoxin

F. Marcucci, D. A. Lappi, M. Ghislieri, D. Martineau, A. Formosa, S. Siena, M. Bregni, M. Soria, A. M. Gianni

Research output: Contribution to journalArticle

Abstract

We have evaluated both the proliferative response as well as the Thy-1 Ag expression of lymphocytes from mice treated in vivo with an anti-Thy-1 immunotoxin (IT). The IT was a rat IgG2c mAb recognizing the Thy-1 Ag, disulfide-linked to a ribosome-inactivating protein isolated from the seeds of the plant Saponaria officinalis (soapwort). Toxicity studies showed that a single i.v. injection of doses up to 20 μg IT/mouse was well tolerated and allowed indefinite survival. The Con A-induced proliferative response of spleen cells from mice killed 1 day after treatment with sublethal doses of IT was inhibited in a dose-dependent manner, with complete inhibition observed at doses of ≥5 μg IT/mouse. Control experiments showed that the inhibition was due to the IT and not its single components. Moreover, the IT effect was abolished by a large (100-fold) excess of anti-Thy-1 mAb alone given concurrently, but not by an unrelated, isotype-matched rat mAb. At all IT doses, the proliferative response to a B cell mitogen (LPS) was normal. Kinetic studies showed a time- and dose-dependent reconstitution of Con A responsiveness. In limiting dilution cultures of spleen cells from mice treated with 5 μg IT 1 or 4 days before death, a 97% depletion of T lymphocytes capable of proliferation was observed. Limiting dilution cultures showed that also the thymus of IT-treated mice was depleted by more than 90% of growth-competent T lymphocytes. Cytofluorographic studies of Thy-1+ cells from the spleens of IT-treated mice gave results which did not correlate with those obtained in functional assays. Thus, a dose-dependent reduction, followed by a time-dependent reconstitution of Thy-1+ cells was observed in this case too, but the depletion occurred at later time points and was less complete than that observed in functional assays. Moreover, the mean fluorescence intensity of the residual Thy-1+ cells decreased below normal levels.

Original languageEnglish
Pages (from-to)2955-2960
Number of pages6
JournalJournal of Immunology
Volume142
Issue number8
Publication statusPublished - 1989

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Immunotoxins
T-Lymphocytes
Spleen
Saponaria
Thomsen-Friedenreich antibodies
Ribosome Inactivating Proteins
Mitogens
Disulfides
Thymus Gland
Seeds
B-Lymphocytes
Cell Culture Techniques
Fluorescence

ASJC Scopus subject areas

  • Immunology

Cite this

Marcucci, F., Lappi, D. A., Ghislieri, M., Martineau, D., Formosa, A., Siena, S., ... Gianni, A. M. (1989). In vivo effects in mice of an anti-T cell immunotoxin. Journal of Immunology, 142(8), 2955-2960.

In vivo effects in mice of an anti-T cell immunotoxin. / Marcucci, F.; Lappi, D. A.; Ghislieri, M.; Martineau, D.; Formosa, A.; Siena, S.; Bregni, M.; Soria, M.; Gianni, A. M.

In: Journal of Immunology, Vol. 142, No. 8, 1989, p. 2955-2960.

Research output: Contribution to journalArticle

Marcucci, F, Lappi, DA, Ghislieri, M, Martineau, D, Formosa, A, Siena, S, Bregni, M, Soria, M & Gianni, AM 1989, 'In vivo effects in mice of an anti-T cell immunotoxin', Journal of Immunology, vol. 142, no. 8, pp. 2955-2960.
Marcucci F, Lappi DA, Ghislieri M, Martineau D, Formosa A, Siena S et al. In vivo effects in mice of an anti-T cell immunotoxin. Journal of Immunology. 1989;142(8):2955-2960.
Marcucci, F. ; Lappi, D. A. ; Ghislieri, M. ; Martineau, D. ; Formosa, A. ; Siena, S. ; Bregni, M. ; Soria, M. ; Gianni, A. M. / In vivo effects in mice of an anti-T cell immunotoxin. In: Journal of Immunology. 1989 ; Vol. 142, No. 8. pp. 2955-2960.
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abstract = "We have evaluated both the proliferative response as well as the Thy-1 Ag expression of lymphocytes from mice treated in vivo with an anti-Thy-1 immunotoxin (IT). The IT was a rat IgG2c mAb recognizing the Thy-1 Ag, disulfide-linked to a ribosome-inactivating protein isolated from the seeds of the plant Saponaria officinalis (soapwort). Toxicity studies showed that a single i.v. injection of doses up to 20 μg IT/mouse was well tolerated and allowed indefinite survival. The Con A-induced proliferative response of spleen cells from mice killed 1 day after treatment with sublethal doses of IT was inhibited in a dose-dependent manner, with complete inhibition observed at doses of ≥5 μg IT/mouse. Control experiments showed that the inhibition was due to the IT and not its single components. Moreover, the IT effect was abolished by a large (100-fold) excess of anti-Thy-1 mAb alone given concurrently, but not by an unrelated, isotype-matched rat mAb. At all IT doses, the proliferative response to a B cell mitogen (LPS) was normal. Kinetic studies showed a time- and dose-dependent reconstitution of Con A responsiveness. In limiting dilution cultures of spleen cells from mice treated with 5 μg IT 1 or 4 days before death, a 97{\%} depletion of T lymphocytes capable of proliferation was observed. Limiting dilution cultures showed that also the thymus of IT-treated mice was depleted by more than 90{\%} of growth-competent T lymphocytes. Cytofluorographic studies of Thy-1+ cells from the spleens of IT-treated mice gave results which did not correlate with those obtained in functional assays. Thus, a dose-dependent reduction, followed by a time-dependent reconstitution of Thy-1+ cells was observed in this case too, but the depletion occurred at later time points and was less complete than that observed in functional assays. Moreover, the mean fluorescence intensity of the residual Thy-1+ cells decreased below normal levels.",
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AU - Marcucci, F.

AU - Lappi, D. A.

AU - Ghislieri, M.

AU - Martineau, D.

AU - Formosa, A.

AU - Siena, S.

AU - Bregni, M.

AU - Soria, M.

AU - Gianni, A. M.

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