Complement Activation Determines the Therapeutic Activity of Rituximab In Vivo

Rituximab is an anti-CD20 chimeric mAb effective for the treatment of B-NHL. It can lyse lymphoma cells in vitro through both C- and Ab-dependent cellular cytotoxicity. The mechanism of action of rituximab in vivo is however still unclear. We have set up a new in vivo model in nonimmunodeficient mic...

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Published inThe Journal of immunology (1950) Vol. 171; no. 3; pp. 1581 - 1587
Main Authors Di Gaetano, Nicola, Cittera, Elena, Nota, Rachele, Vecchi, Annunciata, Grieco, Valeria, Scanziani, Eugenio, Botto, Marina, Introna, Martino, Golay, Josee
Format Journal Article
LanguageEnglish
Published United States Am Assoc Immnol 01.08.2003
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Abstract Rituximab is an anti-CD20 chimeric mAb effective for the treatment of B-NHL. It can lyse lymphoma cells in vitro through both C- and Ab-dependent cellular cytotoxicity. The mechanism of action of rituximab in vivo is however still unclear. We have set up a new in vivo model in nonimmunodeficient mice by stable transduction of the human CD20 cDNA in the murine lymphoma line EL4. Animals injected i.v. with the EL4-CD20(+) lymphoma cells died within 30 days with evident liver, spleen, and bone marrow involvement, confirmed by immunohistochemistry and PCR analysis. A single injection of rituximab or the murine anti-CD20 Ab 1F5, given i.p. 1 day after the tumor, cured 100% of the animals. Indeed, at week 4 after tumor cell inoculation, CD20(+) cells were undetectable in all organs analyzed in rituximab-treated animals, as determined by immunohistochemistry and PCR. Rituximab had no direct effect on tumor growth in vitro. Depletion of either NK cells or neutrophils or both in tumor-injected animals did not affect the therapeutic activity of the drug. Similarly, rituximab was able to eradicate tumor cells in athymic nude mice, suggesting that its activity is T cell independent. In contrast, the protective activity of rituximab or the 1F5 Ab was completely abolished in syngeneic knockout animals lacking C1q, the first component of the classical pathway of C (C1qa(-/-)). These data demonstrate that C activation is fundamental for rituximab therapeutic activity in vivo.
AbstractList Abstract Rituximab is an anti-CD20 chimeric mAb effective for the treatment of B-NHL. It can lyse lymphoma cells in vitro through both C- and Ab-dependent cellular cytotoxicity. The mechanism of action of rituximab in vivo is however still unclear. We have set up a new in vivo model in nonimmunodeficient mice by stable transduction of the human CD20 cDNA in the murine lymphoma line EL4. Animals injected i.v. with the EL4-CD20+ lymphoma cells died within 30 days with evident liver, spleen, and bone marrow involvement, confirmed by immunohistochemistry and PCR analysis. A single injection of rituximab or the murine anti-CD20 Ab 1F5, given i.p. 1 day after the tumor, cured 100% of the animals. Indeed, at week 4 after tumor cell inoculation, CD20+ cells were undetectable in all organs analyzed in rituximab-treated animals, as determined by immunohistochemistry and PCR. Rituximab had no direct effect on tumor growth in vitro. Depletion of either NK cells or neutrophils or both in tumor-injected animals did not affect the therapeutic activity of the drug. Similarly, rituximab was able to eradicate tumor cells in athymic nude mice, suggesting that its activity is T cell independent. In contrast, the protective activity of rituximab or the 1F5 Ab was completely abolished in syngeneic knockout animals lacking C1q, the first component of the classical pathway of C (C1qa−/−). These data demonstrate that C activation is fundamental for rituximab therapeutic activity in vivo.
Rituximab is an anti-CD20 chimeric mAb effective for the treatment of B-NHL. It can lyse lymphoma cells in vitro through both C- and Ab-dependent cellular cytotoxicity. The mechanism of action of rituximab in vivo is however still unclear. We have set up a new in vivo model in nonimmunodeficient mice by stable transduction of the human CD20 cDNA in the murine lymphoma line EL4. Animals injected i.v. with the EL4-CD20 super(+) lymphoma cells died within 30 days with evident liver, spleen, and bone marrow involvement, confirmed by immunohistochemistry and PCR analysis. A single injection of rituximab or the murine anti-CD20 Ab 1F5, given i.p. 1 day after the tumor, cured 100% of the animals. Indeed, at week 4 after tumor cell inoculation, CD20 super(+) cells were undetectable in all organs analyzed in rituximab-treated animals, as determined by immunohistochemistry and PCR. Rituximab had no direct effect on tumor growth in vitro. Depletion of either NK cells or neutrophils or both in tumor-injected animals did not affect the therapeutic activity of the drug. Similarly, rituximab was able to eradicate tumor cells in athymic nude mice, suggesting that its activity is T cell independent. In contrast, the protective activity of rituximab or the 1F5 Ab was completely abolished in syngeneic knockout animals lacking C1q, the first component of the classical pathway of C (C1qa super(-/-)). These data demonstrate that C activation is fundamental for rituximab therapeutic activity in vivo.
Rituximab is an anti-CD20 chimeric mAb effective for the treatment of B-NHL. It can lyse lymphoma cells in vitro through both C- and Ab-dependent cellular cytotoxicity. The mechanism of action of rituximab in vivo is however still unclear. We have set up a new in vivo model in nonimmunodeficient mice by stable transduction of the human CD20 cDNA in the murine lymphoma line EL4. Animals injected i.v. with the EL4-CD20(+) lymphoma cells died within 30 days with evident liver, spleen, and bone marrow involvement, confirmed by immunohistochemistry and PCR analysis. A single injection of rituximab or the murine anti-CD20 Ab 1F5, given i.p. 1 day after the tumor, cured 100% of the animals. Indeed, at week 4 after tumor cell inoculation, CD20(+) cells were undetectable in all organs analyzed in rituximab-treated animals, as determined by immunohistochemistry and PCR. Rituximab had no direct effect on tumor growth in vitro. Depletion of either NK cells or neutrophils or both in tumor-injected animals did not affect the therapeutic activity of the drug. Similarly, rituximab was able to eradicate tumor cells in athymic nude mice, suggesting that its activity is T cell independent. In contrast, the protective activity of rituximab or the 1F5 Ab was completely abolished in syngeneic knockout animals lacking C1q, the first component of the classical pathway of C (C1qa(-/-)). These data demonstrate that C activation is fundamental for rituximab therapeutic activity in vivo.
Author Nota, Rachele
Vecchi, Annunciata
Introna, Martino
Di Gaetano, Nicola
Scanziani, Eugenio
Golay, Josee
Cittera, Elena
Botto, Marina
Grieco, Valeria
Author_xml – sequence: 1
  fullname: Di Gaetano, Nicola
– sequence: 2
  fullname: Cittera, Elena
– sequence: 3
  fullname: Nota, Rachele
– sequence: 4
  fullname: Vecchi, Annunciata
– sequence: 5
  fullname: Grieco, Valeria
– sequence: 6
  fullname: Scanziani, Eugenio
– sequence: 7
  fullname: Botto, Marina
– sequence: 8
  fullname: Introna, Martino
– sequence: 9
  fullname: Golay, Josee
BackLink https://www.ncbi.nlm.nih.gov/pubmed/12874252$$D View this record in MEDLINE/PubMed
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Snippet Rituximab is an anti-CD20 chimeric mAb effective for the treatment of B-NHL. It can lyse lymphoma cells in vitro through both C- and Ab-dependent cellular...
Abstract Rituximab is an anti-CD20 chimeric mAb effective for the treatment of B-NHL. It can lyse lymphoma cells in vitro through both C- and Ab-dependent...
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SubjectTerms Animals
Antibodies, Monoclonal - administration & dosage
Antibodies, Monoclonal - therapeutic use
Antibodies, Monoclonal, Murine-Derived
Antibody-Dependent Cell Cytotoxicity - genetics
Antibody-Dependent Cell Cytotoxicity - immunology
Antineoplastic Agents - administration & dosage
Antineoplastic Agents - therapeutic use
Cell Division - genetics
Cell Division - immunology
Cell Survival - genetics
Cell Survival - immunology
Complement C1q - deficiency
Complement C1q - genetics
Complement C1q - physiology
Complement Pathway, Classical - genetics
Complement Pathway, Classical - immunology
Humans
Lymphocyte Depletion
Lymphoma - immunology
Lymphoma - pathology
Lymphoma - prevention & control
Lymphoma - therapy
Mice
Mice, Inbred C57BL
Mice, Knockout
Mice, Nude
Neoplasm Metastasis - immunology
Neoplasm Metastasis - prevention & control
Neoplasm Metastasis - therapy
Neoplasm Transplantation
Neutrophils - pathology
Rituximab
Tumor Cells, Cultured
Title Complement Activation Determines the Therapeutic Activity of Rituximab In Vivo
URI http://www.jimmunol.org/cgi/content/abstract/171/3/1581
https://www.ncbi.nlm.nih.gov/pubmed/12874252
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