Archives of Dermatological Research

, Volume 264, Issue 2, pp 213–223 | Cite as

A new method for obtaining viable cells from dermal infiltrates

A study on 2,4 dinitrochlorobenzene induced contact dermatitis
  • L. Molnár
  • J. M. Baló-Banga
  • J. Leibinger
  • K. Kiraly
Original Works
  • 18 Downloads

Summary

Experimental contact dermatitis has been induced in 2,4 dinitrochlorobenzene (DNCB) sensitized guinea pigs. The developing dermal infiltrate was excised and the infiltrating cells were obtained by mechanical extraction alone as well as by the combination with collagenase and elastase treatment. The most viable cells appeared in the elastase and mechanically extracted samples and the least in those subjected to mechanical treatment alone. The most cells in the enzyme-treated samples were present 24 h after re-exposure of the sensitized animals to DNCB consisting mainly of lymphocytes and of polymorphonuclear granulocytes. The optimum conditions for the action of enzymes including optimum duration of the treatment, buffer milieu, aspecific proteolytic effect on foreign substrate and action on T and B cell receptors have been elaborated. It was concluded that 80 min of collagenase treatment with gentle mechanical extraction under specified conditions does not affect any measurable immunologic properties of the liberated cells resulting in the second best yield. A comparison of these data with earlier reports and their significance is being discussed.

Key words

Eczema Proteolytic enzymes Dermal cells Receptors 

Zusammenfassung

Das experimentelle Kontaktekzem wurde an mit 2,4 Dinitrochlorbenzol (DNCB) sensibilisierten Meerschweinchen ausgelöst. Das nachfolgende dermale Infiltrat wurde herausgeschnitten und die Infiltratszellen durch ausschließlich mechanische Extraktion sowie auch durch Kombination mit Kollagenase- und Elastase-Behandlung gewonnen. Die größte Menge von lebendigen Zellen wurde in den mechanisch und Elastase behandelten, die geringste in den ausschließlich mechanisch behandelten Mustern gefunden. Die Zahl der Zellen war 24 h nach der Reexposition der mit DNCB sensibilisierten Tieren am höchsten. Der Extrakt bestand vorwiegend aus Lymphocyten und polymorphkernigen Granulocyten. Die optimalen Verhältnisse für die Enzymreaktionen, u.a. Dauer der Behandlung, Pufferkonzentration, nichtspezifische Proteolyse und die Wirkung auf T-Zellen-Rezeptoren wurden ausgearbeitet. Nach den Resultaten der Untersuchungen wurde festgestellt, daß eine 80 min lang dauernde Kollagenase-Behandlung kombiniert mit mechanischer Extraktion unter den beschriebenen Verhältnissen keine meßbare Wirkung auf die immunologischen Eigenschaften der Zellen hat.

Schlüsselwörter

Ekzem Proteolytische Enzyme Dermale Zellen Rezeptoren 

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Copyright information

© Springer-Verlag 1979

Authors and Affiliations

  • L. Molnár
    • 1
  • J. M. Baló-Banga
    • 1
  • J. Leibinger
    • 1
  • K. Kiraly
    • 1
  1. 1.The National Institute of Dermatology and Venereology of HungaryBudapestHungary

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