Bacteriolytic activity of human gingival exudate.

We investigated the bacteriolytic activity of gingival crevicular fluid (CF) on 14C-labeled Streptococcus faecalis, Streptococcus mutans, Staphylococcus aureus, and on whole dental plaque. CF was collected from 100 healthy donors pooled and centrifuged at 200 g. CF supernate and a frozen and thawed extract of the pellet were interacted with the different bacterial strains, while Streptococcus faecalis and Staphylococcus aureus released 60% and 75% of the radioactive label, only 38% of it was solubilized from Streptococcus mutans, following their incubation with the CF supernate. The findings agreed with results obtained by interacting bacteria with a frozen and thawed lysate of human peripheral blood leukocytes. On the other hand, extracts from frozen and thawed CF pellet were inactive. Further, lipoteichoic acid and lipopolysaccharide were released by CF from Gram-positive and Gram-negative bacteria, respectively. The role of bacteriolytic factors, present in CF, as a result of the interaction between microorganisms and leukocytes at inflammatory sites is discussed.

Inflammatory lesions and bone resorption induced in the rat periodontium by lipoteichoic acid of Streptococcus mutans.

Severe inflammatory lesions were induced in the periodontal tissues of the rat following the intragingival injection of lipoteichoic acid (LTA) from Streptococcus mutans. There was no difference in the severity and distribution of the lesions between nonimmunized rats and animals immunized against LTA after antigenic challenge. The lesions are characterized by the occurrence of granulation tissue, massive infiltration of PMNs, abscess formation, bone resorption, and new bone formation. Deacylated LTA and saline caused relatively mild inflammation, and no significant bone resorption or new bone formation was evident. The peak response was reached after 3 intragingival infections. The mechanisms by which LTA caused the pathological alterations in the rat periodontium and the possible relations of this experimental model to periodontal disease in the human are discussed.

Modulation of human lymphocyte transformation by bacterial products and leukocyte lysates.

Blast transformation of human peripheral blood lymphocytes by PHA is shown to be modulated by lipoteichoic acid (LTA) of Streptococcus mutans, by a cell-sensitizing factor of Actinomyces viscosus, as well as by a frozen and thawed extract of human leukocytes (LE). While small amounts of LE (5-50 micrograms/10(6) cells) significantly enhanced PHA-induced transformation, higher amounts showed a lesser effect on the blastogenic response. Both LTA and the A. viscosus extract did not cause any lymphocyte blastogenic effect when used alone. On the other hand LTA had an inhibitory effect and the A. viscosus extract had an enhancing effect when lymphocytes were pretreated by these agents and then exposed to PHA.

Rapid and direct detection of herpes simplex virus and varicella-zoster virus antigens in clinical specimens by staphylococcal reagent and membrane filtration.

Herpes simplex virus was detected rapidly and directly in 31 and varicella-zoster virus in two of 50 clinical specimens using specific antisera, stabilized staphylococci rich in protein A and membrane filtration. Microscopical examination of the cells retained on the filter membrane revealed attached staphylococci only on cells harbouring viral antigens but not on non-infected cells or cells from healthy donors. Infected cells treated with negative control sera and stabilized staphylococci and subsequently subjected to membrane filtration were also devoid of the marker. It was also possible to detect herpes simplex virus antigens in three specimens which were culture negative. Similar results were obtained with staphylococci specifically sensitized with anti-herpes simplex rabbit serum. No interfering, non-specific background of unattached staphylococci was observed on the filter membrane. The results were confirmed by the direct immunofluorescent test. The method is sensitive, specific, and provides results within 3 h. It could be employed for the rapid screening of populations at risk, e.g. pregnant women, medical personnel, and societies with a growing incidence of genital herpes. Since no special, expensive equipment is required, the method is also suitable for modestly equipped clinical laboratories.

Rapid detection of herpes simplex virus and varicella-zoster virus in clinical specimens by the use of Staphylococcus aureus rich in protein A.

Herpes simplex virus (HSV) type 1 and type 2 and varicella-zoster virus (VZV) were detected in cytospin preparations from clinical material by using specific antisera and Staphylococcus aureus, Cowan 1 strain, rich in protein A (SRA). Virus was isolated in tissue culture from 22 of the 30 specimens submitted for examination. Eighteen isolates showed cytopathic effect characteristic of HSV infection and 4 of VZV. In the cytospin preparations of the same samples HSV was detected in 15, two contained too few cells to allow a reliable diagnosis and one sample was negative when the SRA reagent was used. In the cytospin preparations of 2 of the 8 samples, which did not show cytopathic effect on isolation in tissue culture, HSV was detected by the SRA. This points to the possible presence of inactive virus in the specimens. All 4 cases of VZV infection were diagnosed correctly with the staphylococcal reagent. No reaction was observed between VZV antigens and rabbit anti-HSV sera. Cells in which viral infection was detected by specific antisera and SRA did not show staphylococcal adherence to their surface after interaction with normal rabbit or normal human serum. Similarly, cells from healthy donors, treated with positive and negative sera were found negative. The method is easy to perform and results can be obtained within three hours from the time specimens are received at the laboratory. Its use offers a rapid diagnosis in suspected cases of herpetic infections in which early therapy is recommended.

The use of Staphylococcus aureus rich in protein A in the detection of herpes simplex virus antigens.

Herpes simplex virus (HSV) type 1 antigens were detected in infected human embryonic lung cells with the aid of specific antiserum and Staphylococcus aureus rich in protein A. When such staphylococci carrying specific anti-HSV IgG on their surface were interacted with various suspension of virus, a reduction in the initial virus titre of about 65% was obtained. However, no direct coagglutination was observed between cell-free supernatants of HSV or HSV-infected cells and sensitized staphylococci. When monolayers or suspended cells infected with the virus were treated with dilutions of specific anti-HSV antiserum followed by non-sensitized staphylococci (indirect method), an "aureola" of the bacteria was detected around the cells expressing the viral antigens. A similar picture was observed when infected cells were interacted directly with sensitized staphylococci. Viral antigens were detected already 12 hours post infection, well before the appearance of cytopathic effect. The sensitivity of the indirect method was found to be higher than that of the direct one and dependent on the multiplicity of infection and the serum dilution used. The method is proposed as a rapid means of identifying viral antigens in diagnostic and experimental virology.

Excretion of alpha-foetoprotein in the urine of pregnant rats and hepatoma-bearing animals.

Urine of normal rats, pregnant animals and animals bearing chemically induced hepatoma was tested with antisera to foetoproteins by the double immunodiffusion technique. Antigens were not detected in the urine of normal rats. Alpha-foetoprotein was demonstrated in the urine of pregnant rats and hepatomabearing animals.

Comparison and evaluation of fluorescent antibody techniques in the detection of herpes simplex virus in oral infection.

This article deals with the reliability and applicability of fluoroscopy in the diagnosis of herpetic lesions of the oral cavity. The reported results are based on a clinical experiment performed on fifty-two lesions suspected to be of herpetic origin. Three different methods were compared for detection of the possible presence of the virus--cytologic, virologic, and immunofluorescent. The greater reliability and speed of the last technique are demonstrated by our findings. Such features acquire special importance in the differential diagnosis of a number of diseases.

Placental antigens and fetoproteins in the urine of rats - indicators of resorption of conceptuses.

Placenta-specific antigens and alpha-fetoprotein were detected in the urine of rats with induced abnormal pregnancies. Moreover, in those cases, in which the presumptive diagnosis of intrauterine fetal death or resorption was made, urinary excretion of placental antigens persisted for at least 5 days.

Antigenic identity of alpha-macrofetoprotein and acute phase protein in the rat.

Antisera were raised in rabbits against rat embryonic blood and inflammatory serum. The antisera, made specific by absorption procedures, were cross reacted with embryonic blood and inflammatory serum. A reaction of identity between alpha-macrofetoprotein and acute phase protein was demonstrated.

Antibodies in the saliva after antigenic challenge of the parotid gland of systemically sensitized rats.

Anti-BSA antibodies were detected in the parotid saliva of untreated rats. In non-sensitized animals, the parotid duct of which was repeatedly instilled with saline or BSA, the antibody titers rose significantly. Introduction of BSA into the parotid gland of sensitized animals was followed by a significant rise in salivary but not in circulating antibodies. The findings demonstrate the relative independence of the local from the systemic immune system.

Experimental allergic sialoadenitis. VII. Reactivity of the parotid gland to antigenic challenge in passively immunized rats.

Rats were passively immunized by an intraperitoneal injection of homologous anti-BSA serum and their salivary glands were challenged 20 min or 24 hours later with BSA by the intraductal route. Immune complex sialoadenitis developed only when challenge was early. It is concluded that immunoglobulins are transferred from the circulation into the salivary glands and are relatively rapidly cleared by a mechanism yet unknown, possibly by salivary flow.

Excretion of liver antigens in the urine of patients with hepatic diseases.

Liver antigens were detected in the urine of 4 of 42 patients with various liver diseases. The urine of 25 healthy subjects and patients with diseases not affecting the liver was devoid of antigens in detectable amounts. The presence of hepatic antigens in the urine did not correlate with severity of jaundice and SGOT levels but correlated with parenchymal necrosis and was associtated with a high mortaltiy.

Experimental allergic sialoadenitis. IX. The regional lymph nodes after antigenic challenges of the parotid glands in rats.

The regional lymph nodes of the parotid gland of rats were examined histologically. The nodes were removed from untreated animals or rats given instillations of BSA or saline into the parotid duct. Anti-BSA antibodies were assessed in the saliva and serum. Patterns of reactivity of the lymph nodes did not correlate with the presence or absence of salivary and serum antibodies. The occurrence of antibodies in the saliva appears to reflect the activity of immunocytes residing in the parotid gland. The morphological features of the lymph nodes correspond to the overall immunological experience.

Alpha-M-fetoprotein in serum of rats after experimental manipulation in the oral cavity.

Alpha-M-fetoprotein (AMFP) was detected in the serum of rats subjected to intraoral manipulations, i.e., instillations into the parotid gland and injections into the gingival mucosa of saline or foreign proteins. The identity of AMFP with acute-phase protein is pointed out.