Antitoxic Cholera Immunity in Mice: Influence of Antigen Deposition on Antitoxin-Containing Cells and Protective Immunity in Different Parts of the Intestine

Abstract

The importance of the mode of antigen presentation (intravenous, oral, or enteral restricted to the lower ileum) in the development of a local immune response and immunological memory for such a response in different parts of the intestine was studied in mice. Cholera toxin was used as antigen and the immune response was assayed by determining both the number of specific antitoxin-containing cells in the lamina propria and protection against experimental cholera. The results showed that all of these routes of antigen presentation could induce significant memory along the entire small intestine. In contrast, the actual production of antitoxin-containing cells or protective immune response elicited by booster immunization was restricted to those parts of the intestine that were directly exposed to antigen; i.e., lower ileum boosting resulted in immunity in the distal ileum but not in the proximal jejunum, whereas oral or intravenous boosting gave a response in both jejunum and ileum. Protection correlated closely with the number of antitoxin-containing cells in the lamina propria (correlation coefficient, 0.88); ≥4,000 antitoxin-containing cells per mm 3 conferred solid immunity to cholera toxin-induced diarrhea. The total number of immunoglobulin-containing cells in intestines was not significantly influenced by the specific immunizations. There were four times as many of these cells in the upper jejunum (167,000 cells per mm 3 ) as in the lower ileum, but the proportions of immunoglobulin A-containing cells (80 to 85%), immunoglobulin M-containing cells (14 to 20%), and immunoglobulin G-containing cells (0.4 to 0.9%) were similar in various parts of the intestine. The results indicate a differential dependence on local tissue antigen for the intestinal antibody-secreting cells and their memory cell precursors.

A Sensitive Method for the Detection of Specific Antibody Production in Different Isotypes from Single Lamina Propria Plasma Cells

“…Lycke spleen after repeated intravenous immunizations (80% IgG) as opposed to that seen in lamina propria cells after oral immunizations (80% IgA) suggested that the sensitivity for detection of the different isotypes was high. The representation of the various isotypes in the lamina propria antitoxin response also closely resembled that previously reported for total Ig-containing cells using ACC technique [12]. Furthermore.…”

“…The sensitivity of this assay compared with other available techniques for detection of antigen-specific gut mucosal responses appears to be very high. The immunohistochemical detection of antibody-containing cells (ACC) in tissue sections or the measurement of specific antibodies in intestinal washings or in supernatants from cell cultures can only poorly detect antigenspecific responses after a single oral priming immunization with CT, whereas the method presented in this paper registered substantial primary antitoxin SFC responses in the lamina propria, especially when the sensitivity-enhancing vaseline border ring method was used [8,12,13,17]. Furthermore, the lower limit of sensitivity ofthe ACC technique has been reported to be around 100 ACC/mm\ with peak responses of 10,000-20.000 ACC/mm' to oral immunizations with CT [17].…”

“…Despite this progress, the study of antigenspecific responses of the local immune system and the cellular mechanisms responsible for mounting such a response has been hampered by the lack of suitable methods for assessing immunoglobulin secretion at the single cell level. To date, most studies on specific antibody responses by the gut mucosal immune system have been performed with immunohistological methods demonstrating antibody-containing cells (ACC) in tissue sections, or ELISA methods determining specific antibodies in intestinal washings or supernatants from in vitro cultured gut lymphocytes [8,12,13,17]. These methods have limitations In that they do not provide any possibility of studying regulatory influences on individual B lymphocytes of the gut.…”

Effect of allergen immunotherapy on nasal responses in guinea‐pigs with allergic rhinitis

“…Olympus, Tokyojapan) in combination with a light microscope. The number of mast cells per mm" was calculated by Lange's method [14].…”

The Secretory IgA System

“…197 (1985) The Seeretory IgA Syndrom 331 secretory IgA antibodies protect against rhino-, myxo-and paramyxovirus infections (11). In experimental models it has also been clearly shown that secretory IgA antibodies can protect against cholera (27,30,32) and that this protection relates to the number of IgA-producing cells in the small intestine (30,32). In the human it has recently been demonstrated that secretory IgA in milk confers protection of the breastfed baby against Vibrio cholerae (18).…”