Functional assay of the alternative complement pathway of rat serum

Coonrod, J D; Jenkins, Susan D.

doi:10.1016/0022-1759(79)90142-x

Cited by 11 publications

(7 citation statements)

References 13 publications

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“…Bronchoalveolar lavage fluid from [10][11][12] Sprague-Dawley rats (-30 ml) was freed of leukocytes and concentrated 20-fold by filtration through an Amicon membrane (10,000-mol wt exclusion). The concentrate was incubated with washed, log-phase pneumococci at 370C and serial cultures were performed (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Pneumococcal serotypes 1, 3, and 25 (American Type Culture Collection, Rockville, MD) were maintained in an encapsulated state by monthly passage in mice, and stored in heat-inactivated rabbit serum at 40C (12). Dr. Alexander Tomasz (The Rockefeller University, New York) generously provided a pneumococcal "lysis-defective" transformant (R36A pneumococcus transformed with DNA from a lysis-defective mutant) containing <1% of the murein hydrolase activity of the wild strain (13).…”

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confidence: 99%

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Detection and partial characterization of antibacterial factor(s) in alveolar lining material of rats.

Coonrod¹,

Yoneda²

1983

J. Clin. Invest.

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Section: Resultsmentioning

confidence: 99%

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confidence: 99%

Detection and partial characterization of antibacterial factor(s) in alveolar lining material of rats.

Coonrod¹,

Yoneda²

1983

J. Clin. Invest.

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“…Opsono-phagocytosis assay. Opsonization and phagocytosis were evaluated as described previously (6). Briefly, bacteria were radiolabeled with [3H]thymidine, washed, and heat-killed.…”

Section: Methodsmentioning

confidence: 99%

“…Resident alveolar macro-phages of Swiss mice and several strains of rats (6a, 21) have been noted to lack detectable complement receptors, and this finding may explain why these macrophages are unable to ingest complement-coated particles. Exudate cells, however, readily ingest type 25 pneumococci once the organisms have been opsonized with complement (6,7). Other investigators (13) have noted that pneumococci in general are not as easily opsonized in vitro as are S. aureus, but the significance of these differences in terms of phagocytosis in vivo is unknown.…”

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confidence: 99%

Comparative role of complement in pneumococcal and staphylococcal pneumonia

Coonrod¹,

Yoneda²

1982

Infect Immun

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To evaluate the role of complement in pneumococcal and staphylococcal pneumonia, we decomplemented rats with cobra venom factor and inoculated them intratracheally with Staphylococcus aureius or type 25 pneumococci. S. aureus produced a patchy bronchopneumonia in normal Sprague-Dawley or Lewis rats, and decomplementation did not increase the severity of staphylococcal infection in either rat strain as judged by quantitative cultures of the lungs and blood at 6, 24, and 48 h after inoculation. In contrast, decomplementation markedly increased the severity of pneumonia caused by type 25 pneumococci in Sprague-Dawley and Lewis rats. In Sprague-Dawley rats, decomplementation significantly increased the number of bacteria in the lungs at 3, 6, and 24 h of infection. Bacteremia developed early in decomplemented Sprague-Dawley rats, but the higher pulmonary bacterial counts did not appear to be caused by bacteremic seeding of the lungs. Decomplemented Sprague-Dawley rats inoculated intravenously with pneumococci failed to develop the very high levels of bacteria in the lungs that were observed when the rats were inoculated intratracheally. Moreover, decomplemented Lewis rats inoculated intratracheally with pneumococci developed significantly increased numbers of pneumococci in the lungs early in infection (3 and 6 h) when they had no detectable bacteremia. These data indicate that in murine models complement plays a major protective role against type 25 pneumococci in the lung, whereas complement is not important to host defense in staphylococcal pneumonia.

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“…C3 activity was measured by immune adherence (3). Alternative pathway activity was titrated to the 50% endpoint with rabbit erythrocytes (4).…”

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Pulmonary opsonins in Klebsiella pneumoniae pneumonia in rats

Coonrod¹

1981

Infect Immun

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Klebsiella pneumoniae was inoculated intrabronchially into rats, and bronchoalveolar lavage fluid and sera were obtained during the ensuing pneumonia. Klebsiellae recovered by lavage were not maximally coated with C3, as judged by studies with fluorescent antibody, whereas the organisms could be coated fully with C3 by a brief incubation in rat serum. The levels of Cl and C3 in lavage fluid obtained during infection were only a small fraction of the levels in the serum, and klebsiellae were not opsonized during incubation with concentrated lavage fluid. Systemic decomplementation did not affect the severity of K. pneumoniae pneumonia, as judged by the measurement of lung weight or by the numbers of klebsiellae in the lungs, but decomplemented rats had a larger number of klebsiellae in the blood at 24 h of infection than did controls. There were fewer klebsiellae in the lungs 4 h after inoculation of preopsonized organisms than after inoculation of organisms which were incubated in control (heat-inactivated) sera. These studies indicate that the concentration of complement and heat-labile opsonins within the alveoli is lower than that in the systemic circulation and is not adequate for effective opsonization of klebsiellae. Complement has been shown to exert a protective effect in a variety of bacterial infections. The precise role of complement in pulmonary defenses, however, has not been well defined. Normal bronchoalveolar lavage fluid contains functional components of both the classical and the alternative complement pathways (10, 12-14), but there is little information about the levels of these components in the lungs during bacterial infection. To assess the role of complement within the pulmonary alveoli during infection, bronchoalveolar lavage fluids of rats with Klebsiella pneumoniae pneumonia were studied. The course of pneumonia in decomplemented rats and in rats injected with preopsonized klebsiellae was also studied. The results of the studies suggest that complement levels within the alveoli are much lower than the levels in the circulation and are not optimal for opsonization of klebsiellae. MATERIALS AND METHODS Animals. Adult (200to 300-g) male Sprague-Dawley rats (Charles River Breeding Laboratories, Wilmington, Mass.) were used throughout. Experimental pneumonia. K. pneumoniae type

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Functional assay of the alternative complement pathway of rat serum

Cited by 11 publications

References 13 publications

Detection and partial characterization of antibacterial factor(s) in alveolar lining material of rats.

Detection and partial characterization of antibacterial factor(s) in alveolar lining material of rats.

Comparative role of complement in pneumococcal and staphylococcal pneumonia

Pulmonary opsonins in Klebsiella pneumoniae pneumonia in rats

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