In Vivo and In Vitro Anti‐Nuclear Reactions in Lupus Patients

Abstract

HE antinuclear factors found in the T sera of patients with systemic lupus erythematosus (SLE) bind to the nuclei of a variety of mammalian cells as measured by immunofluorescent techniques.l Yet tissues from lupus patients rarely show gamma globulin (presumably antinuclear factors) bound in vivo to cell nuclei.2J Evidence has been presented4 suggesting that tissue nuclei of lupus patients do not react with antinuclear factors in vitro as consistently as do nuclei from normal individuals -possibly implying a lack of available binding sites in the tissues of lupus patients.In this study, the capacity of leukocyte nuclei from lupus patients to react in vitro and in oivo with various types of antinuclear factors was studied, with a comparison of the patterns of nuclear fluorescence induced by certain antinuclear MATERIAL AND METHODSSmears were made from the peripheral blood of normal subjects and of 18 patients with typical SLE (with varying degrees of disease activity). Serum specimens were obtained simultaneously from these individuals. These sera exhibited a variety of nuclear fluorescence patterns (diffuse, peripheral rim, speckled) when incubated with normal leukocyte nuclei in o h . In addition, 2 other serum specimens from lupus patients (patients RS and HM) were used. One serum (RS) had consistently induced "peripheral rim" nuclear fluorescence, and the other serum (HM) had led to a combination of "speckled" and "diffuse" nuclear fluorescence when incubated with leukocytes of normal subjects.Peripheral blood smears were utilized in the direct or indirect fluorescent antibody method of detccting antinuclear factors as advocated by Barnett et al.7 Smears were fixed in 95 per cent ethanol at 37 C for 30 minutes and then dried prior to use, evcept where noted below. RESULTSA peripheral blood smear from each lupus patient was incubated with fluorescein-labelled anti-human gamma globulin goat serum (Hyland Labs) looking for gamma globulin bound in viuo to cell nuclei. In 17 of the 18 smears, no nuclear fluorescence was seen. In one smear, weak nuclear fluorescence was seen in a minority of the leukocytes (Table 1).A serum specimen from each lupus p7-tient was incubated for 30 minutes with an autologous peripheral blood smear and with a normal donor's blood smear. After being washed with phosphate-buffered saine (PBS), the smears were then incubated with fluorescein-labelled anti-human gamma globulin goat serum. Nuclear fluorescence was seen after incubation with 17 of the 18 sera whether autologous or normal donor blood smears were used (Table 1). The nuclear fluorescence pattern seen after incubation of a particular lupus serum with normal blood leukocytes was generally but not always similar to the pattern resulting

Ερευναι Επι Των Αντιπυρηνικων Αντισωματων

“…Ή έμμεσος μέθοδος τοϋ άνοσοφθορισμοΰ επιτρέπει τήν ακριβή έντόπισιν τοϋ σημείου αντιδράσεως τών εν τω ορώ αντισωμάτων μετά τών πυρηνικών στοιχείων. Διά ταύτης αποδεικνύεται, δτι καίτοι τα διάφορα αντιπυρηνικά αντισώματα αντιδρούν έκλεκτικώς μετά διαφόρων συστατικών τοΰ πυρήνος, δεν είναι ειδικά τοΰ είδους, δεδομένου, δτι θετικαί αντιδράσεις δυνατόν να ληφθούν δια χρησιμοποιήσεως υποστρώματος (αντιγόνου) εκ πυρήνων -ορ γάνων διαφόρων ζώων και άνθρωπου ως και περιφερικού α'ίματος (Beck, 1961, Lachmann and Kunkel, 1961, Weir et al, 1961, Zweiman and Hildreth, 1968). Οιονδήποτε δθεν έμπύρηνον κύτταρον δυνατόν να χρησιμοποιηθή ως υπόστρωμα δια τήν άναζήτησιν των αντιπυρηνικών αντισωμάτων υπό την άπαραίτητον προϋπόθεσιν να ύπάρχη βλάβη της κυτταρικής μεμβράνης, 'ίνα καταστή εφικτή ή είσοδος των αντισωμάτων καί ή προσπέλασις τοϋ πυρήνος.…”