Secretion of IL-4 from human basophils: The relationship between IL-4 mRNA and protein in resting and stimulated basophils

Donald MacGlashan, Jane M. White, Shau Ku Huang, Santa J. Ono, John T. Schroeder, Lawrence M. Lichtenstein

Research output: Contribution to journalArticle

231 Citations (Scopus)

Abstract

Purified human basophils have been examined for secretion of IL-4 protein and expression of IL-4 mRNA after stimulation with several secretagogues. In general, these studies used a 15-min preincubation with IL-3, before challenge with secretagogues. Under these conditions, IL-4 release averaged 30 pg/106 basophils (range 4-70) after challenge with anti-IgE Ab. FMLP and C5a led to somewhat lower levels of secretion. A direct comparison of basophils at 88 to 99% purity with basophils from the same preparations, but at lower purities, showed that the amount of IL-4 secretion was proportional to the purity of the basophils. The presence of mRNA for IL-4 (as determined by reverse transcriptase-PCR, competitive reverse transcriptase-PCR, or Northern blots) was also strictly related to the purity of the basophils. IL- 4 mRNA was also found to be constitutively present and was increased after stimulation. The concentration of polyclonal anti-IgE Ab required for optimal IL-4 release was somewhat less than that required for optimal histamine release. IL-4 secretion was slower (t( 1/2 ) of 1.5 hr than histamine release and was inhibited by cycloheximide. In a final series of studies, we found that IL-3 was not required for IL-4 secretion; a short 15-min preincubation with IL-3 resulted in the same or slightly less IL-4 release than no treatment with IL-3. In contrast, an 18-h pretreatment with IL-3 resulted in a nearly tenfold increase in IL-4 secretion. We conclude that human basophils secrete IL-4 in response to several secretagogues and that IL-3 priming is not necessary to observe IL-4 secretion.

Original languageEnglish
Pages (from-to)3006-3016
Number of pages11
JournalJournal of Immunology
Volume152
Issue number6
Publication statusPublished - Mar 15 1994
Externally publishedYes

Fingerprint

Basophils
Interleukin-4
Messenger RNA
Interleukin-3
Proteins
Histamine Release
Reverse Transcriptase Polymerase Chain Reaction
Cycloheximide
Northern Blotting

ASJC Scopus subject areas

  • Immunology

Cite this

MacGlashan, D., White, J. M., Huang, S. K., Ono, S. J., Schroeder, J. T., & Lichtenstein, L. M. (1994). Secretion of IL-4 from human basophils: The relationship between IL-4 mRNA and protein in resting and stimulated basophils. Journal of Immunology, 152(6), 3006-3016.

Secretion of IL-4 from human basophils : The relationship between IL-4 mRNA and protein in resting and stimulated basophils. / MacGlashan, Donald; White, Jane M.; Huang, Shau Ku; Ono, Santa J.; Schroeder, John T.; Lichtenstein, Lawrence M.

In: Journal of Immunology, Vol. 152, No. 6, 15.03.1994, p. 3006-3016.

Research output: Contribution to journalArticle

MacGlashan, D, White, JM, Huang, SK, Ono, SJ, Schroeder, JT & Lichtenstein, LM 1994, 'Secretion of IL-4 from human basophils: The relationship between IL-4 mRNA and protein in resting and stimulated basophils', Journal of Immunology, vol. 152, no. 6, pp. 3006-3016.
MacGlashan, Donald ; White, Jane M. ; Huang, Shau Ku ; Ono, Santa J. ; Schroeder, John T. ; Lichtenstein, Lawrence M. / Secretion of IL-4 from human basophils : The relationship between IL-4 mRNA and protein in resting and stimulated basophils. In: Journal of Immunology. 1994 ; Vol. 152, No. 6. pp. 3006-3016.
@article{bb9bc1a441e247a28ed44fb25a045e53,
title = "Secretion of IL-4 from human basophils: The relationship between IL-4 mRNA and protein in resting and stimulated basophils",
abstract = "Purified human basophils have been examined for secretion of IL-4 protein and expression of IL-4 mRNA after stimulation with several secretagogues. In general, these studies used a 15-min preincubation with IL-3, before challenge with secretagogues. Under these conditions, IL-4 release averaged 30 pg/106 basophils (range 4-70) after challenge with anti-IgE Ab. FMLP and C5a led to somewhat lower levels of secretion. A direct comparison of basophils at 88 to 99{\%} purity with basophils from the same preparations, but at lower purities, showed that the amount of IL-4 secretion was proportional to the purity of the basophils. The presence of mRNA for IL-4 (as determined by reverse transcriptase-PCR, competitive reverse transcriptase-PCR, or Northern blots) was also strictly related to the purity of the basophils. IL- 4 mRNA was also found to be constitutively present and was increased after stimulation. The concentration of polyclonal anti-IgE Ab required for optimal IL-4 release was somewhat less than that required for optimal histamine release. IL-4 secretion was slower (t( 1/2 ) of 1.5 hr than histamine release and was inhibited by cycloheximide. In a final series of studies, we found that IL-3 was not required for IL-4 secretion; a short 15-min preincubation with IL-3 resulted in the same or slightly less IL-4 release than no treatment with IL-3. In contrast, an 18-h pretreatment with IL-3 resulted in a nearly tenfold increase in IL-4 secretion. We conclude that human basophils secrete IL-4 in response to several secretagogues and that IL-3 priming is not necessary to observe IL-4 secretion.",
author = "Donald MacGlashan and White, {Jane M.} and Huang, {Shau Ku} and Ono, {Santa J.} and Schroeder, {John T.} and Lichtenstein, {Lawrence M.}",
year = "1994",
month = "3",
day = "15",
language = "English",
volume = "152",
pages = "3006--3016",
journal = "Journal of Immunology",
issn = "0022-1767",
publisher = "American Association of Immunologists",
number = "6",

}

TY - JOUR

T1 - Secretion of IL-4 from human basophils

T2 - The relationship between IL-4 mRNA and protein in resting and stimulated basophils

AU - MacGlashan, Donald

AU - White, Jane M.

AU - Huang, Shau Ku

AU - Ono, Santa J.

AU - Schroeder, John T.

AU - Lichtenstein, Lawrence M.

PY - 1994/3/15

Y1 - 1994/3/15

N2 - Purified human basophils have been examined for secretion of IL-4 protein and expression of IL-4 mRNA after stimulation with several secretagogues. In general, these studies used a 15-min preincubation with IL-3, before challenge with secretagogues. Under these conditions, IL-4 release averaged 30 pg/106 basophils (range 4-70) after challenge with anti-IgE Ab. FMLP and C5a led to somewhat lower levels of secretion. A direct comparison of basophils at 88 to 99% purity with basophils from the same preparations, but at lower purities, showed that the amount of IL-4 secretion was proportional to the purity of the basophils. The presence of mRNA for IL-4 (as determined by reverse transcriptase-PCR, competitive reverse transcriptase-PCR, or Northern blots) was also strictly related to the purity of the basophils. IL- 4 mRNA was also found to be constitutively present and was increased after stimulation. The concentration of polyclonal anti-IgE Ab required for optimal IL-4 release was somewhat less than that required for optimal histamine release. IL-4 secretion was slower (t( 1/2 ) of 1.5 hr than histamine release and was inhibited by cycloheximide. In a final series of studies, we found that IL-3 was not required for IL-4 secretion; a short 15-min preincubation with IL-3 resulted in the same or slightly less IL-4 release than no treatment with IL-3. In contrast, an 18-h pretreatment with IL-3 resulted in a nearly tenfold increase in IL-4 secretion. We conclude that human basophils secrete IL-4 in response to several secretagogues and that IL-3 priming is not necessary to observe IL-4 secretion.

AB - Purified human basophils have been examined for secretion of IL-4 protein and expression of IL-4 mRNA after stimulation with several secretagogues. In general, these studies used a 15-min preincubation with IL-3, before challenge with secretagogues. Under these conditions, IL-4 release averaged 30 pg/106 basophils (range 4-70) after challenge with anti-IgE Ab. FMLP and C5a led to somewhat lower levels of secretion. A direct comparison of basophils at 88 to 99% purity with basophils from the same preparations, but at lower purities, showed that the amount of IL-4 secretion was proportional to the purity of the basophils. The presence of mRNA for IL-4 (as determined by reverse transcriptase-PCR, competitive reverse transcriptase-PCR, or Northern blots) was also strictly related to the purity of the basophils. IL- 4 mRNA was also found to be constitutively present and was increased after stimulation. The concentration of polyclonal anti-IgE Ab required for optimal IL-4 release was somewhat less than that required for optimal histamine release. IL-4 secretion was slower (t( 1/2 ) of 1.5 hr than histamine release and was inhibited by cycloheximide. In a final series of studies, we found that IL-3 was not required for IL-4 secretion; a short 15-min preincubation with IL-3 resulted in the same or slightly less IL-4 release than no treatment with IL-3. In contrast, an 18-h pretreatment with IL-3 resulted in a nearly tenfold increase in IL-4 secretion. We conclude that human basophils secrete IL-4 in response to several secretagogues and that IL-3 priming is not necessary to observe IL-4 secretion.

UR - http://www.scopus.com/inward/record.url?scp=0028278699&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028278699&partnerID=8YFLogxK

M3 - Article

C2 - 8144899

AN - SCOPUS:0028278699

VL - 152

SP - 3006

EP - 3016

JO - Journal of Immunology

JF - Journal of Immunology

SN - 0022-1767

IS - 6

ER -