Sites of Formation of Immune Globulins and of a Component of C'3

Abstract

Various methods have been used to isolate tissues or cell types from animals in order to determine the sites of plasma protein formation. Studies on the effect of extirpation of (1, 2) and of damage to (3-6) liver and spleen have indicated the role of these organs, but more specific evidence has been obtained with the use of perfused isolated organs (7-9) and of tissue culture (10-19).The results of such studies have shown that the liver is the main, if not the only site of albumin formation; and moreover, that most of the o~-and some of the fl-globulins are formed in the liver. The formation of clotting factors (16) and fibrinogen (7,13) by the liver was also demonstrated; and the studies of Rice el al. (20,21) indicate

The Liver as the Site of C-Reactive Protein Formation

“…Note that labeling of CxRP occurs only in the liver culture. Labeling of ~1c is seen in the spleen and liver cultures as with other species (10)(11)(12)19). The peripheral leukocytes show strong labeling of an al-globulin which has not been further identified.…”

“…Tissue Cultures and Analysis oJ Culture Fluids.--The methods for preparing tissue cultures, media, and handling of culture fluids prior to immunoelectrophoresis have been described in detail elsewhere (10)(11)(12). 100 to 200 mg of minced tissue was cultured in roller tubes for 24 to 48 hr with 2 ml of a medium containing 1 #c of L-isoleucine-C 14 and L-lysine-C 14 each per ml (Institut Pasteur, Paris, France, or Schwarz Bioresearch, Orangeburg, New York) of specific activity 600 to 1000 #c/rag.…”

The Cells of Human Colostrum II. Synthesis of IgA and β1c

“…were cultured with 14 C-labeled amino acids, and the Peripheral blood leukocytes were collected and preradioactive amino acids incorporated into the newly pared for culture as follows. Leukocyte-rich plasma synthesized proteins from the cultures were detected was removed from heparinized blood; it was diluted by immunoelectrophoresis and autoradiography [10]. with 1 volume of HBSS and centrifuged at 300 x g for 5 min.…”

“…Appropriate carriers (unlabeled human serum or colostrum) were added to the antigen wells to facilitate the development of reproducible immunoprecipitin lines [10,25]. Three microliters of the carrier protein and 9 /A of the concentrated culture fluids were added to the antigen wells.…”

Gamma Globulin and Antibody Formation in Vitro

“…The resultant tissue culture fluids were dialyzed against 0.015 M pH 7.2 phosphate-buffered saline, concentrated by lyophilization, and analyzed by means of radioautography of immunoelectrophoretic patterns (16). Since the culture fluids contained only minute amounts of the individual labeled serum proteins, normal rabbit serum was used as a carrier.…”