Microbial etiology of acute pneumonia in hospitalized patients.

The purpose of this study was to determine the microbial etiology of pneumonia by using strict criteria among a group of hospitalized patients. Patients with acute community-acquired or hospital-acquired pneumonia were studied in a systematic and comprehensive manner for bacterial, viral, chlamydial, mycobacterial, and fungal pathogens. A total of 198 patients with 204 episodes of pneumonia were evaluated. Despite 100 percent follow-up of all surviving patients, a specific etiologic agent could be found in only 103 episodes. Among 154 episodes of community-acquired pneumonia, a diagnosis was made in 79; the most common pathogen was from the genus Legionella, followed by various Gram-negative enteric bacteria, Gram-positive cocci, influenza A virus, and Mycoplasma pneumoniae. The etiologic agent was found in 24 of the 50 patients with hospital-acquired pneumonia; no pathogen predominated. We conclude that even when elaborate diagnostic studies are done, including many invasive procedures, the etiology can be determined in only about half of the patients with acute pneumonia. The pathogens of pneumonia in this study are not markedly different between community-acquired and hospital-acquired infection.

Response of guinea pigs to intravaginal inoculation with guinea pig cytomegalovirus.

Female guinea pigs were inoculated intravaginally with guinea pig cytomegalovirus (GPCMV) propagated either in guinea pig embryo fibroblast cultures (GPEF) or salivary glands. The incidence of infection was higher with GPEF virus. Rare instances of isolation of GPCMV from cervical swabs 9-48 hr after inoculation was attributed to survival of inoculum in the genital tract. Neither immunofluorescence microscopy nor histopathologic examination showed evidence for active infection of genital tract tissue examined up to day 5 after inoculation. At necropsy on days 30-49, GPCMV was isolated from salivary glands and occasionally from pancreas and lymph nodes. Seroconversion following intravaginal inoculation was demonstrated by an enzyme-linked immunosorbent assay (ELISA) test, and titers were comparable to those after intraperitoneal or subcutaneous inoculation. However, titers of neutralizing antibody, determined by plaque-reduction assay, were significantly lower in the group inoculated intravaginally. These findings are relevant to consideration of cytomegalovirus as a sexually transmitted agent in humans.

Chlamydial salpingitis in female guinea pigs receiving oral contraceptives.

Female guinea pigs were given daily doses of a combination of oral contraceptive (OC) agents, consisting of mestranol and norethynodrel suspended in sesame oil or distilled H2O, and were infected in the genital tract with the chlamydial agent of guinea pig inclusion conjunctivitis (GPIC). Counts of chlamydial inclusions in cells of vaginal smears collected during infection, showed prolongation and enhancement of infection in OC-treated animals as compared with controls. Appearance of IgG and IgA antibodies to GPIC in genital secretions, as determined by enzyme-linked immunosorbent assay (ELISA), was also delayed in OC-treated animals as compared with controls. OC-treated infected animals were killed on days 15 and 43, and gross pathological evidence for ascending infection culminating in salpingitis was found in all of five and four of five animals, respectively. On the other hand, among untreated infected controls on each sacrifice day, only one of five animals had any evidence for ascending infection. Chlamydiae were detected by light and electron microscopy in fallopian tube tissue collected on day 15 following OC-treatment but not in tissue from control animals.

Specific effect of estradiol on the genital mucosal antibody response in chlamydial ocular and genital infections.

Estradiol treatment of female guinea pigs was found to alter the course of genital, but not ocular, infection with the chlamydial agent of guinea pig inclusion conjunctivitis. Immunoglobulin G (IgG) and IgA responses in genital secretions of genitally infected animals were delayed by estradiol treatment, but neither response in the eye resulting from either ocular or genital infection was affected. However, the appearance of IgG in the genital tract after ocular infection was markedly inhibited in estradiol-treated guinea pigs.

Absence of progesterone effects on chlamydial genital infection in female guinea pigs.

The effect of progesterone alone and in combination with estradiol was investigated in ovariectomized and gonadally intact female guinea pigs infected with the chlamydial agent of guinea pig inclusion conjunctivitis (GPIC). The course of the infection, as determined by the percentage of cells with GPIC (chlamydia) inclusions in Giemsa-stained vaginal scrapings, was not affected in animals receiving 5.0 mg of progesterone daily. Progesterone had no influence on the enhancement of infection by estradiol. In comparison with sesame oil-treated controls, infection was prolonged by four to six days (P less than .05) in animals receiving a combination of 5.0 mg of progesterone plus 1.0 microgram of estradiol or 1.0 microgram of estradiol alone each day. In ovariectomized animals, estradiol delayed the appearance of IgA antibody in genital secretions, whereas progesterone alone had no effect. Guinea pigs treated with estradiol or progesterone plus estradiol manifested an acute endometritis not observed in animals treated with progesterone alone or in controls receiving sesame oil. Although cervical ectopy, analogous to that seen in women with high levels of progesterone, was identified by histopathology in animals treated with progesterone, no enhancement of the chlamydial infection was observed.

Electron microscopic observations concerning the in vivo uptake and release of the agent of guinea-pig inclusion conjunctivitis (Chlamydia psittaci) in guinea-pig exocervix.

This report details electron-microscopical observations concerning C. psittaci infection in vivo. The model employed was that of the guinea-pig infected at the exocervical region with the agent of guinea-pig inclusion conjunctivitis (GPIC). Our observations indicate that chlamydial particles gain access to their target cells by the mechanism of endocytosis. Single GPIC elementary bodies were seen to be positioned within individual endosomes. The observations reported here provide evidence that chlamydial particles that had undergone their developmental cycle within the exocervical epithelial cells may leave the epithelium in 2 ways; within entire infected cells that had been shed into the lumen of the cervix and by means of the liberation of chlamydial particles from disrupted cells. The mechanism of cell disruption and shedding is thought to involve the large number of PMNs observed to be present within the enlarged intercellular spaces of the infected epithelium.

Chronic chlamydial genital infection in congenitally athymic nude mice.

Congenitally athymic nude mice and their heterozygous counterparts were inoculated intravaginally with the chlamydial agent of mouse pneumonitis, a Chlamydia trachomatis biovar. Heterozygous mice resolved their infections in 20 days, whereas nude mice developed chronic infections which lasted at least 265 days and did not resolve within the time course of the experiments. Heterozygous mice produced high levels of antibody in both serum and secretions in contrast to nude mice, which produced very low levels of antibody in serum alone.

Chlamydial pneumonitis induced in newborn guinea pigs.

One- to three-day-old guinea pigs were inoculated intranasally with the chlamydial agent of guinea pig inclusion conjunctivitis. Physical signs of infection included a marked increase in respiration rate on days 5 to 10 of infection and radiographic evidence of pneumonia on day 6. When animals were killed at various times after infection and lung tissue was examined by histopathology, evidence of pneumonia was found beginning on day 4 and lasting as long as day 12, with maximal pathological changes on days 6 to 8. The pneumonia was generally unilateral and consisted of an acute inflammatory component in the bronchioles with granulocytes in both the lumen and the wall of the bronchioles and an interstitial and intra-alveolar mononuclear infiltrate in the parenchyma of the lung. Chlamydial antigen was detected in the bronchial epithelial cells by immunoperoxidase staining, and the guinea pig inclusion conjunctivitis organism was isolated from lung tissue on days 6 to 9. No other significant bacteria were isolated from lung tissue or seen on gram stains of lung sections. Both immunoglobulin M and immunoglobulin G serum antibodies to the guinea pig inclusion conjunctivitis agent were detected as early as day 8 and reached peak levels on day 12. The infection was apparently self-limiting. This model presents the opportunity to investigate pathophysiological and immunological aspects of chlamydial respiratory infections in a neonatal animal.

Effects of various doses of estradiol on chlamydial genital infection in ovariectomized guinea pigs.

The effect of various doses of estradiol on genital tract infection by the chlamydial agent of guinea pig inclusion conjunctivitis (GPIC) was investigated in ovariectomized guinea pigs. Prolongation of infection, as determined by chlamydial inclusion counts of cells in Giemsa-stained smears of vaginal scrapings, was observed in animals receiving daily doses of 1.0, 10.0, 100.0, or 1000 micrograms of estradiol. In contrast to controls, ascending infection resulting in endometritis was found in animals receiving doses of greater than or equal to 1.0 microgram of estradiol per day. Response to estradiol treatment was reflected in an increase in cervical-uterine wet weight and uterine wall thickness. No differences were observed in time of appearance of antibody titers to GPIC in serum, but a delay in appearance of IgA antibody to GPIC in genital secretions was found in estradiol-treated animals receiving doses of greater than or equal to 1.0 microgram per day.

Association of adenovirus type 16 with Reye's-syndrome-like illness and pneumonia.

Thirty-eight antigenically distinct viruses have been described as adenoviruses, though only about one third have been commonly associated with human illnesses. In mid-January 1981, adenovirus 16, one of the more rarely encountered ones, was isolated from three patients in separate hospitals in the Little Rock, Arkansas, area, even though it had not previously been isolated there. We report the details of these cases, which included a Reye's-syndrome-like illness and fatal viral pneumonia. The reports indicate a new and more important clinical role for adenovirus 16 infection.

Immune response in mice infected in the genital tract with mouse pneumonitis agent (Chlamydia trachomatis biovar).

Female Swiss-Webster mice were inoculated intravaginally with mouse pneumonitis agent (MoPn), a Chlamydia trachomatis biovar. Inoculation with 3.5 X 10(5) egg lethal doses per mouse resulted in shedding of the agent from the genital tract for as long as 21 days. Immunoglobulin M antibodies to MoPn were detected in plasma by day 7 post-inoculation, and immunoglobulin G antibodies were detected by day 14. Antibodies were detected in genital secretions by day 20, and titers in plasma and secretions were still considerable on day 56. Delayed-type hypersensitivity tests, determined by footpad swelling, were not positive in appreciable numbers of animals until after day 25. Delayed-type hypersensitivity reactions were maximal 24 h after testing and were preceded by an Arthus-like reaction, which appeared within 3 h and declined by 12 h. Convalescent animals were rechallenged by intravaginal inoculation and were found to be solidly immune.

Effect of antithymocyte serum on the course of chlamydial genital infection in female guinea pigs.

The treatment of female guinea pigs, infected in the genital tract with the chlamydial agent of guinea pig inclusion conjunctivitis, with rabbit anti-guinea pig thymocyte serum extended the course of the infection by 20 to 30 days. The rabbit anti-guinea pig thymocyte serum was shown to suppress delayed hypersensitivity responses to the guinea pig inclusion conjunctivitis agent and the contact allergen oxazolone. The appearance of antibody in genital secretions was delayed, but the infection persisted at low levels even when normal serum and secretory antibody titers were attained, indicating that cell-mediated immunity may play a role in the resolution of chlamydial genital infections.

Humoral immune response in acquired immunity to chlamydial genital infection of female guinea pigs.

Immunity to reinfection in the genital tract of female guinea pigs with the agent of guinea pig inclusion conjunctivitis was found to be dependent upon an intact humoral immune response. Cell-mediated immunity in the absence of humoral immunity had no apparent role in resistance to challenge infection.

Effect of estradiol on chlamydial genital infection of female guinea pigs.

Female guinea pigs were treated daily with 1 mg of beta-estradiol-3-benzoate intramuscularly beginning 14 days before intravaginal inoculation with the chlamydial agent of guinea pig inclusion conjunctivitis and continuing during the course of the infection. Treatment with estradiol was found to markedly influence the course of genital infection with the chlamydial agent of guinea pig inclusion conjunctivitis, producing infections of greater intensity and longer duration than those in control animals. Moreover, pathogenesis was altered in that ascending infection was observed, resulting in endometritis, cystic salpingitis, and cystitis. Infection in the controls was limited to the cervix and vagina. Estradiol treatment increased the apparent number of infected cells in the cervix and vagina as detected by histopathology and immunofluorescent staining. Humoral and cell-mediated immune responses to the chlamydial agent of guinea pig inclusion conjunctivitis were comparable in estradiol-treated and untreated animals. These data indicate that hormonal manipulation may have profound effects on the course of chlamydial genital infections.

Cystitis associated with chlamydial infection of the genital tract in male guinea pigs.

Male guinea pigs were infected with the chlamydial agent of guinea pig inclusion conjunctivitis (GPIC) by intraurethral injection of chlamydiae or by placement of a drop of chlamydial suspension on the meatus of the extruded penis. Transient urethritis and cystitis were observed in animals infected by either method. The production of cystitis by the drop-on technique indicated that chlamydiae are able to ascend the urethra and that the bladder may be a target organ of chlamydial infection. When infected animals were immunosuppressed with cyclophosphamide, the number of guinea pigs with cystitis was increased, and chlamydiae could be detected in the bladder for as long as 50 days after infection. In contrast, GPIC was not detected in the bladders of untreated animals after day 18.

A new animal model for the study of Chlamydia trachomatis genital infections: infection of mice with the agent of mouse pneumonitis.

A new animal model for the study of genital infections caused by Chlamydia trachomatis has been developed. Female mice were successfully infected after intravaginal inoculation with the C. trachomatis agent of mouse pneumonitis. Evidence for infection was obtained by detection of chlamydial inclusions in smears of cervical scrapings treated with Giemsa stain. Chlamydiae were observed in sections of cervical tissues examined by light and electron microscopy as well as by immunofluorescence microscopy. An antibody response to the agent of mouse pneumonitis was also demonstrated in sera after infection. The mouse model of genital infection with the agent of mouse pneumonitis offers an opportunity to investigate many questions related to pathogenesis and immunity associated with C. trachomatis genital infections.

Influenza type A virus infection of suckling mice pre-exposed to insecticide carrier.

Suckling CD-1 outbred mice exposed topically to insecticide carrier (IC), a mixture of emulsifiers and solvent, were rendered less sensitive to infection with lethal doses of influenza type A/PR8/34 (H0N1) virus than untreated and mock-treated control mice. Decreased sensitivity to influenza type A/PR8/34 virus infection was evidenced by a significant increase in the mean percent survival of the mice. In addition, a 10- to 100-fold reduction in the 50% lethal titer of the stock virus was observed in IC-treated mice relative to untreated mice. Decreased sensitivity was virus dose related and occurred within a dose range of 2 to 8 X LD50. No decrease in mortality rate was observed as a function of exposure to IC.

Humoral immunity in the resolution of genital infection in female guinea pigs infected with the agent of guinea pig inclusion conjunctivitis.

Female guinea pigs infected in the genital tract with the chlamydial agent of guinea pig inclusion conjunctivitis were selectively immunosuppressed by varying regimens of cyclophosphamide (Cy) treatment. Temporary suppression of both humoral and cell-mediated immunity by daily treatment of Cy (25 mg/kg) for 13 days resulted in a prolonged infection, whereas daily treatment for the duration of the experiment totally prevented the development of humoral and cell-mediated responses and produced an intense and prolonged infection which did not resolve. When humoral immunity alone was suppressed by treatment with Cy (250 and 150 mg/kg) at 9-day intervals, the infection again did not resolve. Treatment with 100 mg of Cy per kg at 9-day intervals resulted in an extended infection which resolved concomitantly with the development of antibody to guinea pig inclusion conjunctivitis. These data indicate that humoral immunity is essential for the recovery of female guinea pigs from guinea pig inclusion conjunctivitis genital infection. A market weight loss was observed which could not be attributed to Cy treatment alone. Edematous and ulcerative changes of the external genitalia were also noted.