Enhanced binding of peripheral blood mononuclear leukocytes to γ-interferon-treated cultured keratinocytes

B. J. Nickoloff, David Lewinsohn, E. C. Butcher

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Dermatophatologists observe mononuclear leukocytes in close apposition to keratinocytes (KCs) in graft versus host disease and in other lymphocyte-mediated skin diseases, such as lichen planus, erythema multiforme, and lupus erythematosus. Since the KCs are Class II histocompatibility antigen (HLA-DR) positive in these diseases (indicating local production of gamma interferon, IFN-γ, by activated T-cells), we sought to determine whether IFN-γ treatment of KCs would influence the ability of allogeneic peripheral blood mononuclear leukocytes (PBMLs) to adhere to cultured KCs in vitro. The adherence of PBMLs to KC monolayers was determined by the three following methods: (a) methanol fixation of the washed KCs (after PBML incubation), followed by hematoxylin-eosin staining and direct counting of adherent PBMLs; (b) fluorescein isothyiocyanate (FITC) labeling of PBML, followed by measuring the amount of FITC-PBML bound to KCs after washing either by direct visualization with a fluorescence microscope; or by (c) quantitative fluorescence spectroscopy following lysis of the adherent cells. While untreated KCs bound allogeneic PBMLs minimally 15-120 min at 37°C, pretreatment of the KCs with IFN-γ (300 U/ml, 3 days) produced significantly increased binding of the PBMLs by approximately fivefold. By contrast, IFN-α and IFN-β (103 U/ml) had no effect. Also, despite the induction of HLA-DR on cultured human fibroblasts, no increased binding of PBMLs after IFN-γ treatment was observed. The selective ability of IFN-γ to produce a marked increase in adherence between KCs and PBMLs suggests a new role for IFN-γ in the immunobiology of the skin.

Original languageEnglish (US)
Pages (from-to)413-418
Number of pages6
JournalAmerican Journal of Dermatopathology
Volume9
Issue number5
StatePublished - 1987
Externally publishedYes

Fingerprint

Mononuclear Leukocytes
Keratinocytes
Interferon-alpha
HLA-DR Antigens
Fluorescein
Erythema Multiforme
Lichen Planus
Fluorescence Spectrometry
Histocompatibility Antigens Class II
Graft vs Host Disease
Hematoxylin
Eosine Yellowish-(YS)
Skin Diseases
Interferon-gamma
Methanol
Fibroblasts
Fluorescence

ASJC Scopus subject areas

  • Dermatology
  • Pathology and Forensic Medicine

Cite this

Enhanced binding of peripheral blood mononuclear leukocytes to γ-interferon-treated cultured keratinocytes. / Nickoloff, B. J.; Lewinsohn, David; Butcher, E. C.

In: American Journal of Dermatopathology, Vol. 9, No. 5, 1987, p. 413-418.

Research output: Contribution to journalArticle

@article{17f42916dedb4390a946f64135b2ddcf,
title = "Enhanced binding of peripheral blood mononuclear leukocytes to γ-interferon-treated cultured keratinocytes",
abstract = "Dermatophatologists observe mononuclear leukocytes in close apposition to keratinocytes (KCs) in graft versus host disease and in other lymphocyte-mediated skin diseases, such as lichen planus, erythema multiforme, and lupus erythematosus. Since the KCs are Class II histocompatibility antigen (HLA-DR) positive in these diseases (indicating local production of gamma interferon, IFN-γ, by activated T-cells), we sought to determine whether IFN-γ treatment of KCs would influence the ability of allogeneic peripheral blood mononuclear leukocytes (PBMLs) to adhere to cultured KCs in vitro. The adherence of PBMLs to KC monolayers was determined by the three following methods: (a) methanol fixation of the washed KCs (after PBML incubation), followed by hematoxylin-eosin staining and direct counting of adherent PBMLs; (b) fluorescein isothyiocyanate (FITC) labeling of PBML, followed by measuring the amount of FITC-PBML bound to KCs after washing either by direct visualization with a fluorescence microscope; or by (c) quantitative fluorescence spectroscopy following lysis of the adherent cells. While untreated KCs bound allogeneic PBMLs minimally 15-120 min at 37°C, pretreatment of the KCs with IFN-γ (300 U/ml, 3 days) produced significantly increased binding of the PBMLs by approximately fivefold. By contrast, IFN-α and IFN-β (103 U/ml) had no effect. Also, despite the induction of HLA-DR on cultured human fibroblasts, no increased binding of PBMLs after IFN-γ treatment was observed. The selective ability of IFN-γ to produce a marked increase in adherence between KCs and PBMLs suggests a new role for IFN-γ in the immunobiology of the skin.",
author = "Nickoloff, {B. J.} and David Lewinsohn and Butcher, {E. C.}",
year = "1987",
language = "English (US)",
volume = "9",
pages = "413--418",
journal = "American Journal of Dermatopathology",
issn = "0193-1091",
publisher = "Lippincott Williams and Wilkins",
number = "5",

}

TY - JOUR

T1 - Enhanced binding of peripheral blood mononuclear leukocytes to γ-interferon-treated cultured keratinocytes

AU - Nickoloff, B. J.

AU - Lewinsohn, David

AU - Butcher, E. C.

PY - 1987

Y1 - 1987

N2 - Dermatophatologists observe mononuclear leukocytes in close apposition to keratinocytes (KCs) in graft versus host disease and in other lymphocyte-mediated skin diseases, such as lichen planus, erythema multiforme, and lupus erythematosus. Since the KCs are Class II histocompatibility antigen (HLA-DR) positive in these diseases (indicating local production of gamma interferon, IFN-γ, by activated T-cells), we sought to determine whether IFN-γ treatment of KCs would influence the ability of allogeneic peripheral blood mononuclear leukocytes (PBMLs) to adhere to cultured KCs in vitro. The adherence of PBMLs to KC monolayers was determined by the three following methods: (a) methanol fixation of the washed KCs (after PBML incubation), followed by hematoxylin-eosin staining and direct counting of adherent PBMLs; (b) fluorescein isothyiocyanate (FITC) labeling of PBML, followed by measuring the amount of FITC-PBML bound to KCs after washing either by direct visualization with a fluorescence microscope; or by (c) quantitative fluorescence spectroscopy following lysis of the adherent cells. While untreated KCs bound allogeneic PBMLs minimally 15-120 min at 37°C, pretreatment of the KCs with IFN-γ (300 U/ml, 3 days) produced significantly increased binding of the PBMLs by approximately fivefold. By contrast, IFN-α and IFN-β (103 U/ml) had no effect. Also, despite the induction of HLA-DR on cultured human fibroblasts, no increased binding of PBMLs after IFN-γ treatment was observed. The selective ability of IFN-γ to produce a marked increase in adherence between KCs and PBMLs suggests a new role for IFN-γ in the immunobiology of the skin.

AB - Dermatophatologists observe mononuclear leukocytes in close apposition to keratinocytes (KCs) in graft versus host disease and in other lymphocyte-mediated skin diseases, such as lichen planus, erythema multiforme, and lupus erythematosus. Since the KCs are Class II histocompatibility antigen (HLA-DR) positive in these diseases (indicating local production of gamma interferon, IFN-γ, by activated T-cells), we sought to determine whether IFN-γ treatment of KCs would influence the ability of allogeneic peripheral blood mononuclear leukocytes (PBMLs) to adhere to cultured KCs in vitro. The adherence of PBMLs to KC monolayers was determined by the three following methods: (a) methanol fixation of the washed KCs (after PBML incubation), followed by hematoxylin-eosin staining and direct counting of adherent PBMLs; (b) fluorescein isothyiocyanate (FITC) labeling of PBML, followed by measuring the amount of FITC-PBML bound to KCs after washing either by direct visualization with a fluorescence microscope; or by (c) quantitative fluorescence spectroscopy following lysis of the adherent cells. While untreated KCs bound allogeneic PBMLs minimally 15-120 min at 37°C, pretreatment of the KCs with IFN-γ (300 U/ml, 3 days) produced significantly increased binding of the PBMLs by approximately fivefold. By contrast, IFN-α and IFN-β (103 U/ml) had no effect. Also, despite the induction of HLA-DR on cultured human fibroblasts, no increased binding of PBMLs after IFN-γ treatment was observed. The selective ability of IFN-γ to produce a marked increase in adherence between KCs and PBMLs suggests a new role for IFN-γ in the immunobiology of the skin.

UR - http://www.scopus.com/inward/record.url?scp=0023261809&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023261809&partnerID=8YFLogxK

M3 - Article

C2 - 2446518

AN - SCOPUS:0023261809

VL - 9

SP - 413

EP - 418

JO - American Journal of Dermatopathology

JF - American Journal of Dermatopathology

SN - 0193-1091

IS - 5

ER -