✍ Scribed by H. -Ch. Gerber; F. Hörchner; B. Zander
No coin nor oath required. For personal study only.
The irdluence of humoral and secretory antibodies as well as eell supplements on Nippostrongylus brasiliensls was tested in vitro. Adult Sprague-D~wley-rats approximately 12 weeks of age were used in these experiments.
For the in vitro tests the following culture media were used: 25% chicken-embryoextract, sodium casein, pig liver extract and rat serum for the larval stages. The medium for the eultures of adult ST. brasiliensis eonsisted of 5% yeast extract, 15% casein, 30% Krebs-Ringer-solution and 50 % rat serum.
Secretory antibodies were isol~ted from the rinsing fluid of the rat intestines by high pressure filtration (10 to 15 micron), then cleaning of the fluid through a Sephadex G 15 eolumn and finally narrowing down through eollodene capsules. Mast cells were isolated from the peritoncal cavity by Ficoll-gradient-centrifugation.
Various test series were eonducted with the addition of serum er secretory antibodies of repeatedly infected and immune rats to the medium. In these tests there was never a differenee in the influence on growth ner a higher mortality rate of larval er adult N. brasiliensis in contrast to cultures where serum and sceretory antibodies of non-infec~ed animals were used.
A 100% degranulation of mast cel]s from infeeted rats oecurred already within 14 to 22 hours in the cultures of adult N. brasiliensi~. Variations were not noticed in the influence on the viability of N. brasiliensis kept in media for 10 days without er with cell supplements as well as sera of infeeted er not infected rats.
The results from our experiments demonstrated that therc was no variation in the influence on the development and a higher mortality rate of the larval stages and adult Nippo-strongylu8 in media containing either sera and secretory antibodies of infected er not infected rats.
Zusamrnen/assung. Für die Untersuchungen über den Einfluß von humoralen und sekretorisehen Antikörpern sowie Zellzusätzen auf in vitro gehaltene ~Tippostrongylus brasiliensis verwendeten wir Sprague-Dawley-l~atten im Alter von mindestens 12 Wochen. Die für die in vitro-Haltung von N. brasiliensis verwendeten Nährmedien bestanden bei den Larvenkultnren aus je 25% Kükenembryoextrakt, Natriumcaseinat, Schweineleberextrakt und Rattenserum; bei den Adultenkulturen aus einem Gemisch von ¥east Extrakt (5%), Caseinat (15%), Krebs-Ringerlösung (30%) und Rattenserum (50%).
Sekretorische Antikörper wurden aus der Spülflüssigkeit von Rattendärmen nach Über* druekfiltration (10--15 ~), Reinigung über Sephadex G 15 und Einengung durch Kollodiumhülsen gewonnen. Die Mastzellen stammten aus dem Bauchhöhlenexsudat nach Separierung über Ficollzentrifugation.
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