LFA-1 is sufficient in mediating neutrophil transmigration in Mac-1 knockout mice

Huifang Lu, C. W. Smith, B. J. Hughs, D. C. Bullard, J. L. Perrard, C. Ballantvne

Research output: Contribution to journalArticle

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Abstract

LFA-1 (CD11a) and Mac-1 (CD11b) are two major β2 integrins expressed on neutrophils. They mediate neutrophil adhesion and transmigration through endothelium and extracellular matrix. To investigate the specific contributions of Mac-1 and LFA-1 to neutrophil functions in vivo, we developed CD lib-deficient mice. A replacement construct was developed from 129 sv genomic clones which deleted exon 4. Homozygous mice for the Mac-1 mutation (Mac-1-/-) were confirmed completely deficient in Mac-1 by RNA protection assays and flow cytometry with mAb M1/70 (anti-CD11b). Isolated neutrophils from Mac-1-/- mice completely lacked several Mac-1-dependent functions in vitro: They did not (1) adhere to KLH-coated glass; (2) develop oxidative burst when stimulated by serum-activated zymosan particles; and (3) exhibit homotypic aggregation when stimulated by PMA. LFA-1 expression in Mac-1-/- mice was not different from that in Mac-1+/+ siblings, and functioned normally as shown by homotypic aggregation of purified T cells, previously shown to be LFA-1- and ICAM-I-dependent in Mac-P +. When challenged by thioglycolate i.p. for 4 hours, Mac-1-/- mice had similar levels of neutrophil accumulation in the peritoneal cavity with no significant difference from Mac-1+/+ mice: Mac-1+/+ 15.3±5.3x106, Mac-1' I9.6±11.5×106. Systemic treatment with mAb KBA (anti-LFA-1) caused a maximum of 80% inhibition of neutrophil accumulation in Mac-1-/- mice and 60% inhibition in Mac-1+/+ mice. These data demonstrated that in the absence of Mac-1, LFA-1 is sufficient to mediate significant neutrophil transmigration. LFA-1 may play a more important role in neutrophil trafficking than Mac-1.

Original languageEnglish (US)
JournalFASEB Journal
Volume10
Issue number6
StatePublished - 1996

Fingerprint

Lymphocyte Function-Associated Antigen-1
Knockout Mice
neutrophils
Neutrophils
mice
Agglomeration
Thioglycolates
Zymosan
T-cells
Flow cytometry
zymosan
Respiratory Burst
lymphocyte function-associated antigen-1
Peritoneal Cavity
Integrins
integrins
endothelium
extracellular matrix
Exons
Assays

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Lu, H., Smith, C. W., Hughs, B. J., Bullard, D. C., Perrard, J. L., & Ballantvne, C. (1996). LFA-1 is sufficient in mediating neutrophil transmigration in Mac-1 knockout mice. FASEB Journal, 10(6).

LFA-1 is sufficient in mediating neutrophil transmigration in Mac-1 knockout mice. / Lu, Huifang; Smith, C. W.; Hughs, B. J.; Bullard, D. C.; Perrard, J. L.; Ballantvne, C.

In: FASEB Journal, Vol. 10, No. 6, 1996.

Research output: Contribution to journalArticle

Lu, H, Smith, CW, Hughs, BJ, Bullard, DC, Perrard, JL & Ballantvne, C 1996, 'LFA-1 is sufficient in mediating neutrophil transmigration in Mac-1 knockout mice', FASEB Journal, vol. 10, no. 6.
Lu H, Smith CW, Hughs BJ, Bullard DC, Perrard JL, Ballantvne C. LFA-1 is sufficient in mediating neutrophil transmigration in Mac-1 knockout mice. FASEB Journal. 1996;10(6).
Lu, Huifang ; Smith, C. W. ; Hughs, B. J. ; Bullard, D. C. ; Perrard, J. L. ; Ballantvne, C. / LFA-1 is sufficient in mediating neutrophil transmigration in Mac-1 knockout mice. In: FASEB Journal. 1996 ; Vol. 10, No. 6.
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abstract = "LFA-1 (CD11a) and Mac-1 (CD11b) are two major β2 integrins expressed on neutrophils. They mediate neutrophil adhesion and transmigration through endothelium and extracellular matrix. To investigate the specific contributions of Mac-1 and LFA-1 to neutrophil functions in vivo, we developed CD lib-deficient mice. A replacement construct was developed from 129 sv genomic clones which deleted exon 4. Homozygous mice for the Mac-1 mutation (Mac-1-/-) were confirmed completely deficient in Mac-1 by RNA protection assays and flow cytometry with mAb M1/70 (anti-CD11b). Isolated neutrophils from Mac-1-/- mice completely lacked several Mac-1-dependent functions in vitro: They did not (1) adhere to KLH-coated glass; (2) develop oxidative burst when stimulated by serum-activated zymosan particles; and (3) exhibit homotypic aggregation when stimulated by PMA. LFA-1 expression in Mac-1-/- mice was not different from that in Mac-1+/+ siblings, and functioned normally as shown by homotypic aggregation of purified T cells, previously shown to be LFA-1- and ICAM-I-dependent in Mac-P +. When challenged by thioglycolate i.p. for 4 hours, Mac-1-/- mice had similar levels of neutrophil accumulation in the peritoneal cavity with no significant difference from Mac-1+/+ mice: Mac-1+/+ 15.3±5.3x106, Mac-1' I9.6±11.5×106. Systemic treatment with mAb KBA (anti-LFA-1) caused a maximum of 80{\%} inhibition of neutrophil accumulation in Mac-1-/- mice and 60{\%} inhibition in Mac-1+/+ mice. These data demonstrated that in the absence of Mac-1, LFA-1 is sufficient to mediate significant neutrophil transmigration. LFA-1 may play a more important role in neutrophil trafficking than Mac-1.",
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AB - LFA-1 (CD11a) and Mac-1 (CD11b) are two major β2 integrins expressed on neutrophils. They mediate neutrophil adhesion and transmigration through endothelium and extracellular matrix. To investigate the specific contributions of Mac-1 and LFA-1 to neutrophil functions in vivo, we developed CD lib-deficient mice. A replacement construct was developed from 129 sv genomic clones which deleted exon 4. Homozygous mice for the Mac-1 mutation (Mac-1-/-) were confirmed completely deficient in Mac-1 by RNA protection assays and flow cytometry with mAb M1/70 (anti-CD11b). Isolated neutrophils from Mac-1-/- mice completely lacked several Mac-1-dependent functions in vitro: They did not (1) adhere to KLH-coated glass; (2) develop oxidative burst when stimulated by serum-activated zymosan particles; and (3) exhibit homotypic aggregation when stimulated by PMA. LFA-1 expression in Mac-1-/- mice was not different from that in Mac-1+/+ siblings, and functioned normally as shown by homotypic aggregation of purified T cells, previously shown to be LFA-1- and ICAM-I-dependent in Mac-P +. When challenged by thioglycolate i.p. for 4 hours, Mac-1-/- mice had similar levels of neutrophil accumulation in the peritoneal cavity with no significant difference from Mac-1+/+ mice: Mac-1+/+ 15.3±5.3x106, Mac-1' I9.6±11.5×106. Systemic treatment with mAb KBA (anti-LFA-1) caused a maximum of 80% inhibition of neutrophil accumulation in Mac-1-/- mice and 60% inhibition in Mac-1+/+ mice. These data demonstrated that in the absence of Mac-1, LFA-1 is sufficient to mediate significant neutrophil transmigration. LFA-1 may play a more important role in neutrophil trafficking than Mac-1.

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