Phagocytic cell death leads to enhanced release of pro-inflammatory S100A12 in familial Mediterranean fever.

BACKGROUND: Familial Mediterranean fever (FMF) is a prototypical autoinflammatory syndrome associated with phagocytic cell activation. Pyrin mutations are the genetic basis of this disease, and its expression has been shown in monocytes, granulocytes, dendritic cells, and synovial fibroblasts. Pyrin functions as a cytosolic pattern recognition receptor and forms a distinct pyrin inflammasome. The phagocyte-specific protein S100A12 is predominantly expressed in granulocytes and belongs to the group of damage associated molecular patterns (DAMP). S100A12 can be detected at massively elevated levels in the serum of FMF patients, even in clinically inactive disease. Whether this is crucial for FMF pathogenesis is as yet unknown, and we therefore investigated the mechanisms of S100A12 release from granulocytes of FMF patients presenting clinically inactive. RESULTS: We demonstrate that FMF neutrophils from patients in clinical inactive disease possess an intrinsic activity leading to cell death even in exogenously unstimulated neutrophils. Cell death resembles NETosis and is dependent on ROS and pore forming protein gasdermin D (GSDMD), as inhibitors for both are capable of completely block cell death and S100A12 release. When pyrin-activator TcdA (Clostridium difficile toxin A) is used to stimulate, neutrophilic cell death and S100A12 release are significantly enhanced in neutrophils from FMF patients compared to neutrophils from HC. CONCLUSIONS: We are able to demonstrate that activation threshold of neutrophils from inactive FMF patients is decreased, most likely by pre-activated pyrin. FMF neutrophils present with intrinsically higher ROS production, when cultured ex vivo. This higher baseline ROS activity leads to increased GSDMD cleavage and subsequent release of, e.g., S100A12, and to increased cell death with features of NETosis and pyroptosis. We show for the first time that cell death pathways in neutrophils of inactive FMF patients are easily triggered and lead to ROS- and GSDMD-dependent activation mechanisms and possibly pathology. This could be therapeutically addressed by blocking ROS or GSDMD cleavage to decrease inflammatory outbreaks when becoming highly active.

Determination of TNF alpha, interferon alpha, interleukin 2 and reactivity in the leucocyte migration inhibition test in breast cancer patients.

To date, the results concerning the prognostic importance of parameters of cell-mediated immunity in breast cancer patients are very contradictory; moreover, in most of them the results are hardly comparable due to methodological differences and heterogeneous groups of patients. In 123 patients with nonmetastatic breast carcinoma TNF alpha, INF alpha, IL 2 and reactivity in the leucocyte migration inhibition test (LMI-Test) against autologous tumor tissue were determined and the results correlated with the clinical course of the disease up to a maximum of 108 months. In breast cancer patients TNF alpha-serum levels were significantly (p less than 0.05) elevated compared to healthy controls. We also found that patients with progressive disease had higher levels than patients without recurrences. There were no differences concerning the IL-2 and IFN alpha serum levels between cancer patients and controls, nor did we find a correlation with the clinical course of the disease. In 38% of all breast cancer patients examined, a MIF production against tumor tissue could be demonstrated in the LMI-test. There was no difference concerning the LMI-reactivity between the groups of lymph-node negative and positive patients, but the observation that those patients with an unfavourable clinical course respond more frequently with an enhanced macrophage migration and rarely with migration inhibition was considered of notable prognostic significance. According to these results, it is possible that determination of TNF alpha and delayed type hypersensitivity reaction against tumor tissue in the LMI-test is of clinical value for the determination of risk groups.

A Mobile Laparoscopy Unit.

Our experience has led us to the construction of a Mobile Laparoscopy Unit. This unit increases safety and permits the adequate performance of laparoscopy. The Mobile Laparoscopy Unit is an alternative to a special laparoscopy room.

[The importance of herpes simplex virus type 2 (HSV-2) infection pregnancy and the period following birth (author's transl)]. / Die Bedeutung der Herpes-simplex-Virus-Typ 2-(HSV-2)-Infektion für Schwangerschaft und Neugeborenenperiode.

The Type 2 herpes simplex virus is mainly localized in the genital region and is transmitted by sexual intercourse. A distinction must be made between primary and recurrent infection. During pregnancy, infection of the generative tract with HSV-2 can be expected in 0.1% of cases, depending on the patient's social status. It was possible to culture HSV-2 from the cervical secretion in 0.65% of an investigated group of pregnant women. Fifty percent of all infections in pregnancy are asymptomatic. Miscarriages are common before the 20th week of pregnancy (33%), while later there is a slight increase in premature confinement. Where HSV-2 infection exists at the time of birth, cesarean section is the method of choice for delivery. If rupture of the amnion has occurred over 4 hours previously surgical delivery to prevent infection is no longer justified. A start has been made on therapy for herpes genitalis and neonatal herpes, although sufficient experience for therapeutic recommendations has not yet been gathered.

[The technic of "open pelviscopy". A new method for increased safety in laparoscopy]. / Technik der "Offenen Pelviskopie" Eine neue Methode zur Erhöhung der Sicherheit bei der Bauchspiegelung.

The method of "open pelviscopy" is described and compared with the common performance of gynecological pelviscopy. The "open pelviscopy" combines the advantages of laparotomy--preparation of anatomical layers under view of the operator--with those of pelviscopy--minimal traumatization and optimal view at the intraabdominal organs. This method avoids blind puncture of the abdominal cavity, which otherwise is necessary to perform the pneumoperitoneum and to use the optic instruments. Apart from the well established indications for diagnostic and therapeutic gynecological pelviscopy we prefer this method especially in obese patients and in those who formerly had several laparotomies.

[Cervix carcinoma as an immunological model. Part 2. In vitro studies and HLA system typing]. / Das Zervixkarzinom als tumorimmunologisches Modell. Teil 2: In vitro-Untersuchungen und Typisierungen im HLA-System.

Carcinoma of the cervix uteri is one of the most important tumors in women. The tumor induces an immunological defense reaction against itself in the tumor host. The round cell infiltrate at the tumor invasion line, which predominantly consists of small lymphocytes is the morphological equivalent of this interrelationship. In the lymph-nodes of the drainage area of the tumor certain patterns of reaction can be differentiated, which may be interpreted as humoral or cellular immune reactions. Cellular immune reactions in lymph-nodes of patients operated according to the method of Wertheim-Meigs were correlated with a better prognosis of the tumor disease. The hypothesis of an involvement of the HSV-type II in the genesis of the cervical carcinoma is supported by the increased incidence of antibodies against the virus. Therewith, relations between the stage of disease and the age of the patient were found as a possible basis for the virus hypothesis and the immune reaction to the tumor associated antigens an altered genetic situation may be presumed. Typing in the HLA-system demonstrates an accumulation of the frequency of the antigen B12 in patients with carcinoma of the cervix. These investigations may lead to a differentiated outlook with regard to the detection of risk groups and therapeutical consequences for cervical cancer.

[Cell mediated immunity in cervical cancer patients: evaluation by in vitro leukocyte-migration-inhibition-assay (author's transl)]. / Zellulär vermittelte Immunreaktionen bei Patientinnen mit einem Zervixkarzinom: In vitro-Untersuchungen mit dem Leukozyten-Migration-Inhibitions-Test.

Cellular mediated immune reactions (CMI) against tumour associated antigens are the demonstration of an interrelationship between tumour and tumour host. They are related to the stage and prognosis of the disease. 41 patients with cervical cancer were tested by leukocyte-migration-inhibition test. Cryostat sections of 10 micron of the tumour-tissue, gained by excision or curettage and histologically defined, were taken as tumour antigens. The test was performed in Sykes-Moore-Chamber as so-called one-way-test with the lymphocytes/leukocytes and macrophagen of the tumour patient. Reactions against autologous and homologous tumour tissue and against recall antigens (tuberculin and varidase) were tested. There was no overall diminution of cellular mediated immune reactions. CMI was demonstrated in 69.7% (inhibition in 30.3%, stimulation in 39.4%) in the autologous and in 51.3% in the homologous system (21.6% inhibition, 29.7% stimulation).

Evidence that caspase-13 is not a human but a bovine gene.

Caspase-13 was reported to be a member of the human caspase family of proteases (Humke, E., et al., J. Biol. Chem. 273, 15702-15707, 1998). By contrast, a recent study (Lin, X., et al., J. Biol. Chem. 275, 39920-39926, 2000) could not confirm caspase-13 expression in human tissues. When we searched the GenBank database we found several expressed sequence tags (ESTs) from bos taurus completely matching the published caspase-13 sequence. Reverse transcription polymerase chain reaction (RT-PCR) analysis revealed that bovine but not human peripheral blood mononuclear cells express caspase-13. From these cells we cloned two bovine caspase-13 splice variants and found that the sequence of the larger variant was identical to the mRNA published by Humke et al. Our findings strongly suggest that the previously published caspase-13 sequence is not of human origin but represents a bovine gene.

[Cold-lymphocytotoxic antibodies in the sera of cervix-carcinoma patients]. / Kälte-lymphozytotoxische Antikörper (CoCoCy) in Seren von Cervix-Karzinom-Patientinnen (CCa).

Sera of 120 patients with cervical cancer were screened for cold reacting, complement dependant autolympho-cytotoxins (CoCoCy). These antibodies occurred in 21,7% of the patients in comparison with 11,2% of 116 controls. The highest percentage of CoCoCy-antibodies was detected in the sera of patients with cervical cancer stage I (27,6%). Thus demonstration of CoCoCy-antibodies may be of pathogenetic rather than of diagnostic interest. CoCoCy-antibodies could be seen to reflect autoimmune reactions in cervical cancer.

[HL-A polymorphism and its relation to cervix neoplasms]. / Der HLA-Polymorphismus und seine Beziehung zur Zervix-Neoplasie

Twenty-three HLA antigens were investigated in 121 patients with clinical cancer as well as 188 male and 212 female controls. Patients with cervical carcinoma showed significant differences in some HLA frequencies from that of controls. Patients with cervical carcinoma showed significant differences in some HLA frequencies from that of controls. In the localized form of the neophasia HLA-A3 was decreased, HLA-A9 and B 12 were increased. In the disseminated form of cervical carcinoma HLA-A 1 were found decreased and AW 32 and B 12 increased. HLA antigens could be factors, together with others yet unknown, i.e. immune response genes, HSV-2-infection, which produce an increased susceptibility to the disease.

[Seroepidemiological studies related to the association of genital herpes to cervical cancer (author's transl)]. / Seroepidemiologische Untersuchungen zur Frage des Kausalzusammenhangs zwischen der Herpes-genitalis-Infektion und der Entstehung des Kollum-karzinoms

Blood sera of 290 patients were examined for the presence of antibodies against herpes simplex virus type 1 and type 2 (HSV-1 and HSV-2): 1. lues - seronegative individuals, 2. patients with specific lues antibodies, and 3. patients with so-called carcinoma in situ and invasive carcinoma of the uterine cervix. The group of syphilis-seropositive individuals served as control for the group with cervical lesions, because in both groups certain socioepidemiologic factors (promiscuity, early sexual intercourse) appear to be prevalent. The average value for HSV-2-antibodies was significantly higher in patients with cervical lesions than that in lues-seropositive individuals. Also, the average value was higher in the lues-seropositive group than that in the lues-seronegative group. The frequency of HSV-2-antibodies was highest in the group of patients with so-called carcinoma in situ and invasive carcinoma, less in the lues-seropositive group, and even more less in the lues-seronegative group. These results make it unlikely that the connection between cervical carcinoma and HSV-2-infection is a common or "nonsense"-correlation. They suggest a more narrow correlation between both diseases, but leave open whether it consists of either a causal relationship between herpes infection and the development of cervical carcinoma, or a predilection for herpes infection in the cervical region by the carcinomatous tissue.

[Investigation of HL-antigens in cervical cancer patients (author's transl)]. / Untersuchungen zur Bedeutung der Histokompatibilitätsantigene bei Zervixkarzinompatientinnen

Twenty-three of HL-antigens were investigated in 119 patients with clinical cancer and 230 female controls. Patients with cervical carcioma showed significant differences of some HLA frequencies from that of the controls. HLA-A 3 and A 28 were increased, HLA-A 1 was decreased. HL-antigens could be factors, together with others yet unknown, which produce an increased susceptibility to the disease.

Occurrence and characterization of different types of cytotoxic antibodies in pregnant women in relation to parity and gestational age.

Lymphocytotoxic antibodies in sera from 55 pregnant women (of whom 23 were in their first, 21 were in their second, and 11 were in their third pregnancy), as detected by four different microcytotoxicity tests, were predominantly cold reactive and of the IgM class. During pregnancy, there is an increase in lymphocytotoxic antibody formation which is most pronounced after delivery. It is impressive that during first and second pregnancies antibody frequency is similar in the first and second trimesters, whereas in a third pregnancy there is a decrease intermittently. The behavior of antibody frequency in the second trimester may suggest a possible absorption by fetal and/or placental structures. The antibodies seem to be part of a natural immunization process and may play a protective role in pregnancy.

[Lymphocytotoxic antibodies in cervical cancer]. / Lymphozytotoxische Antikörper bei Patientinnen mit einem Zervixkarzinom.

Sera of 120 patients, suffering from cervical cancer (CCa) of different clinical stages (stage 0: n = 27; stage I: n = 29; stage II--IV: n = 64) as well as the sera of 116 healthy individuals as control group were examined for lymphocytoxic antibodies by microlymphocytotoxicity tests, variated in incubation temperatures (15 degress -- 22 degrees -- 37 degrees C) and incubation times (30--180 minutes). At an incubation temperature of 15 degrees C cold reacting lymphocytotoxic antibodies were detected in 26 patients' sera (21.7%) and in 13 control sera (11.2%). At an incubation temperature of 22 degrees C 36 CCa-sera (30%) were positive in comparison to 11 sera of the control group (9.5%). At an incubation 37 degrees C 40 CCa-sera (33.3%) and 7 control sera (6%) were positive. Relating these results to the clinical stages of the carcinoma, the highest percentage of lymphocytotoxicity was found in the sera of patients with an early stage of the disease. Increased evidence of lymphocytotoxic antibodies in the sera of patients with cervical carcinoma supports the assumption, that various immune reactions may be involved in this disease.

[Humoral immune response--cervical cancer patients: lymphocytotoxic antibodies (author's transl)]. / Humorale Immunreaktionen bei Patientinnen mit einem Zervixkarzinom: Lymphozytotoxische Antikörper.

Sera of 120 cervical cancer patients as well as that of 116 healthy control persons were screened for lymphocytotoxic antibodies using microlymphocytotoxic tests variated in temperature and incubation time. In 76 of 120 cancer-sera (63.3%) lymphocytotoxic antibodies could be detected in comparison with 26 sera of 116 controls (22.4%); p less than 0.0005. With the differentiation into the stages of the disease the highest percentage of lymphocytotoxic antibodies was found in patients with stage 0 of cervical cancer. The presence of lymphocytotoxic antibodies in sera of cervical cancer patients suggest the possibility that this disease is an allotransplant as well as expression of an autoimmune process.

[Anaesthesia and immunology (author's transl)]. / Anästhesie und Immunologie: Eine Ubersicht.

There is evidence of immunosuppressive influence of trauma/operation and anesthesia. The human immunological system is outlined as a complex functioning apparatus with humoral and cellular factors. The influence of operation and/or anesthesia on immunocompetence has been demonstrated by animal studies, in vitro examinations and clinical investigations. The analysis of these results and their significance for an impairment of the immunological surveillance of the patient is tried to be elucidated. There should be payed attention to the clinical consequences, especially in patients on high risk.