Biomedical subjects

C R Rossi

Publications and source records attributed to C R Rossi.

At least 55 records · Page 3Linked to original sources

PubMed1992

Characterization and activation requirements of bovine lymphocytes acquiring cytotoxic activity after interleukin-2 treatment.

Interleukin-2 (IL-2) treatment of cells and generation of non-major histocompatibility complex (MHC)-restricted cytotoxic cells from peripheral blood mononuclear leukocytes (PBML) was studied. Effector-target conjugate assays demonstrated that bovine PBML bound but did not lyse K562, HL60S and HL60R cells unless activated with IL-2. The magnitude of IL-2-activated killing of tumor cells as well as the magnitude of antibody-dependent cellular cytotoxicity depended on the IL-2 concentration. A short treatment (12-18 h) of effector cells with IL-2 was sufficient for development of cytotoxic activity. Withdrawal of IL-2 from the culture resulted in a reduction of cytotoxic activity that could be restored by further addition of IL-2. Cytotoxic activity of IL-2-activated populations obtained after nylon wool or Sephadex G-10 passage, and Percoll gradient centrifugation of PBML suggests that lymphokine-activated killer (LAK) cell activity in PBML is mainly mediated by a non-adherent lymphocyte lacking markers for B-cells. Positive and negative selection experiments using cell sorting confirmed these findings and demonstrated that the cell responsible for LAK cell activity in cattle belongs to a non-monocyte, non-B, CD2+ lymphocyte population. Furthermore, cytotoxic activity could not be generated in CD2+ populations enriched for cells expressing molecules equivalent to human and murine CD4 and CD8. These findings suggest that effector cells mediating non MHC-restricted cytotoxicity in cattle prevail in a population bearing a CD2+, CD4-, CD8- phenotype and that this population depends on the continuous presence of IL-2 for optimal cytotoxic function.

Animals↗

PubMed1992

Adriamycin in hyperthermic perfusion for advanced limb sarcomas.

From February 1989 to April 1991, 12 hyperthermic limb perfusions (HLP) with adriamycin (ADR) were performed in 12 patients with high grade soft tissue sarcomas (9 in the leg and 3 in the arm); two were at A.J.C. stage IIb, eight stage IIIb and two stage IV. ADR (0.7-1.5 mg/kg) was administered in bolus at a mean temperature of 41.5 degrees C and perfused for 60 min. No systemic toxicity was reported; seven patients had grade II locoregional toxicity, another four grade III and one grade IV. Tumor necrosis (radiological evaluation) was less than 25% in two patients, 25% to 50% in two, 50% to 75% in five and greater than 75% in three. Limb sparing surgery was feasible in ten patients. At present eight patients are alive; three had local recurrence and four distant metastases.

Adult↗

PubMed1992

[A comparison between echography and computed tomography in assessing neoplastic recurrences in superficial soft tissues].

Thirty-seven superficial soft-tissue recurrences were evaluated with ultrasonography (US) and computed tomography (CT) to assess the correct diagnostic approach. US and CT examinations were performed at the same time. High-frequency US probes and a third-generation CT scanner were employed; all the lesions underwent also histology or cytology. US correctly identified as recurrences or fibrous tissue all the 37 lesions, whereas CT diagnosed 30 lesions only. Seven of the 14 recurrences < 2 cm diameter were not demonstrated. In conclusion, US provides more reliable information than CT relative to small lesions, which suggests that US must be performed just after therapy. Nevertheless, when bone involvement is suspected, CT is required and its use is also suggested to monitor distant metastases.

Biopsy, Needle↗

PubMed1991

Chemically induced viral resistance in single preimplantation bovine embryos.

Viral mRNA encoding vesicular stomatitis virus glycoprotein sequences was detected and quantitated using a DNA-hybridization dot-blot technique. This assay was employed to determine if the synthetic double-stranded polynucleotide complex of polyriboinosinic-polyribocytidylic acid would elicit viral resistance in vitro in single day 9 bovine embryos. The levels of viral mRNA were assayed in 4 groups of bovine embryos: unexposed, virus-exposed, polynucleotide-treated, and virus plus polynucleotide treated. Reduced quantities of viral mRNA in single polynucleotide treated embryos demonstrated that resistance to viral infection was induced in day 9 bovine embryos.

Animals↗

PubMed1989

Laparoscopy in the abdominal staging of melanoma.

A retrospective study on laparoscopic results in the abdominal staging of 297 patients with melanoma, is reported. In 195 (99.3%) of patients, laparoscopy was technically feasible, and in 10% the clinical stage was changed on the basis of the findings. In 34% of patients with metastasis of the liver and peritoneum, the tumor had a diameter of under 1 cm and therefore was not detectable with non-invasive techniques. In the light of these results laparoscopy is advisable in patients with Stage I melanoma at a high risk of metastatization (Clark IV-V; Breslow greater than 0.76 mm) and in Stage II patients. In Stage III patients, laparoscopy seems indicated for an evaluation of the efficacy of systemic treatment.

Adolescent↗

PubMed1988

The sensitivity of domestic animal cell lines to eight recombinant human interferons.

The biological activities of eight recombinant human interferons (IFNs) were tested on permanent cell lines and low-passage embryonic cultures from domestic animal tissues. Each titer was compared to its titer on Madin-Darby bovine kidney (MDBK) cells. Although different cells within a species had different sensitivities to different IFNs, the order of sensitivity, in general, was ox, sheep, pig, cat, horse, and dog, the latter being quite insensitive to most IFNs.

Animals↗

PubMed1988

Acetaldehyde oxidation in rat liver mitochondria, action of Mg2+, ATP and rotenone on acetaldehyde oxidation in intact mitochondria, and on some purified aldehyde dehydrogenase isozymes.

In intact rat liver mitochondria acetaldehyde is oxidized by three functionally distinct dehydrogenase systems. Two of these reduce intramitochondrial nicotinamide adenine dinucleotide (NAD): one is operative with micromolar acetaldehyde concentrations and is stimulated by Mg2+, the other is operative with millimolar acetaldehyde concentrations and is stimulated by adenosine 5'-triphosphate (ATP). The third system reduces added NAD and is stimulated by rotenone. Connected to these systems, three aldehyde dehydrogenase isozymes (ALDH) have been purified: a low-Km ALDH activated by Mg2+, a high-Km ALDH activated by ATP and Mg2+, a high-Km ALDH activated by rotenone. The properties of some isozymes are affected by detergents. Thus, deoxycholate augments the stimulation of low-Km isozyme by Mg2+ and confers sensitivity to Mg2+ and ATP on one of the high-Km isozymes. A fourth isozyme has been purified. Its affinity for acetaldehyde is so low that it is very unlikely that acetaldehyde is the physiological substrate.

Acetaldehyde↗

PubMed1987

Cytotoxic capabilities of bovine lymphocytes after lectin stimulation.

Bovine peripheral blood mononuclear leukocytes (PBML) were stimulated in vitro with the mitogenic lectins concanavalin A (Con A) and phytohemagglutinin. Their cytotoxic capabilities were evaluated in a 51Cr release assay. Lectin-activated bovine effector cells did not mediate antibody dependent cellular cytotoxicity (ADCC) nor direct killing against cultured tumor target cells. Nevertheless, activation of PBML with lectins consistently generated effector cells able to mediate lectin-dependent cellular cytotoxicity. Cultivation of Con A stimulated-PBML for 3 to 4 weeks in the presence of lymphokines-containing IL-2 generated cells with the ability to mediate lysis without using Con A-coated target cells. However, cytotoxic cultures capable of mediating direct lysis of target cells were not able to mediate ADCC.

Animals↗

PubMed1987

Natural cell-mediated cytotoxicity to cells infected with infectious bovine rhinotracheitis virus.

Cell-mediated cytotoxicity against viral-infected cells was demonstrated in a 6-hr 51Cr release assay. Peripheral blood mononuclear leukocytes from both infectious bovine rhinotracheitis virus (IBRV)-infected and noninfected cattle exhibited preferential lysis against IBRV-infected primary bovine embryonic kidney (BEK) cells compared to cells infected with pseudorabies virus and noninfected BEK cells. Addition of specific antibody to the assay did not enhance cytotoxicity. The effector cell was a nonadherent cell which was either spontaneously enriched or generated during in vitro cultivation. Maximal cytotoxic activity was detected in peripheral blood mononuclear cells cultured for 3 to 5 days. Several factors affected the magnitude of cytotoxicity during the assay: target cell type, concentration of viral inoculum, duration of effector and target cell contact. It is suggested that target cell lysis was a form of natural cell-mediated cytotoxicity mediated by a cell which has different characteristics from the typical human and murine NK cell.

Animals↗

PubMed1986

A monoclonal antibody that identifies mature T lymphocytes of cattle.

Hybridomas to bovine leukocytes were produced by immunization of BALB/C mice with bovine lymphoblasts and fusion of the mouse spleen cells with mouse myeloma cells. Monoclonal antibodies (MABs) were tested against various cell populations by indirect fluorescent microscopy using fluorochrome conjugated antibodies to mouse immunoglobulins. MAB-15, one of the resulting MABs obtained after cloning antibody-producing hybridomas, reacted with 56.8 +/- 8.4% of peripheral blood mononuclear cells (PBMC). MAB-15 did not react with monocytes or B cells, but did react with T cells (fluorescein isothiocyanate-conjugated peanut agglutinin positive cells). MAB-15 reacted with 3.2% of thymocytes from adult cattle. In addition to reacting with T cells, MAB-15 reacted with neutrophils and eosinophils. MAB-15 was characterized as an IgM antibody that was unable to lyse PBMC in the presence of complement. Thus, MAB-15 is a useful marker of mature T cells in the mononuclear cell population.

Animals↗

PubMed1986

Cell-mediated cytotoxicity of peripheral blood mononuclear cells stimulated in vitro for infectious bovine rhinotracheitis virus-infected cells.

Peripheral blood mononuclear cells (PBMC) from calves infected with and hyperimmunized to infectious bovine rhinotracheitis virus (IBRV) were stimulated in vitro with viral antigens to evaluate their cytotoxicity for a variety of cells. The 51-Cr release assay was used to measure cytotoxicity. Cytotoxicity was not present in fresh nonstimulated cells, but was detected in cultured, IBRV-stimulated cells at day 3, was maximal at day 7, and declined thereafter. PBMC stimulated in vitro with IBRV expressed a preference for killing IBRV-infected cells compared to pseudorabies virus (PRV)-infected cells. IBRV-infected, but not PRV-infected, cold target cells inhibited lysis of IBRV-labeled target cells. High concentrations of IBRV hyperimmune serum partially blocked cytotoxicity. Cells expressing a viral preference for cytotoxicity showed no preference for lysis of autologous compared to heterologous bovine cells. PBMC from calves that were either IBRV-immune or not immune were cultured without IBRV stimulation and had similar levels of cytotoxicity for IBRV-infected cells as cells from IBRV-infected cattle.

Animals↗

PubMed1986

In vitro induction of cytotoxic lymphocytes from infectious bovine rhinotracheitis virus hyperimmune cattle.

Cytotoxic lymphocytes against infectious bovine rhinotracheitis virus (IBRV)-infected cells were induced by in vitro stimulation with IBRV antigen of peripheral blood leukocytes obtained from hyperimmune cattle. After one in vitro stimulation with IBRV, cytotoxicity was detected against IBRV-infected autologous target cells, but not against heterologous IBRV-infected or K562 target cells. After 4 in vitro IBRV stimulations at weekly intervals (in the presence of autologous feeder cells followed by interleukin 2 treatment at 2-day intervals), cytotoxicity was detected against autologous-, and to a lesser extent, heterologous-infected cells. Most cells in cytotoxic cultures were positive to a monoclonal antibody shown to react with bovine T lymphocytes.

Animals↗

PubMed1986

Cytotoxicity of bovine lymphocytes after treatment with lymphokines.

Cytotoxic lymphocytes were generated from bovine peripheral blood mononuclear leukocytes after in vitro stimulation with lymphokines that contained interleukin-2. Lymphokine-stimulated cultures were cytotoxic to K562 cells (human natural killer [NK] targets) and YAC-1 cells (mouse NK targets), but not to HSB-2 cells (human NK targets) in a 4-hour, 51Cr-release assay. Cells generated after lymphokine activation also mediated antibody-dependent cellular cytotoxicity to HSB-2 cells. Appearance of effector cells as a function of time in culture, method of stimulation, and cold target competition experiments strongly indicated that direct cytotoxicity and antibody-dependent cellular cytotoxicity may have been mediated by the same cell. Cells generated by similar conditions were able to mediate cytotoxicity against infectious bovine rhinotracheitis virus-infected target cells, especially in an 18-hour assay.

Animals↗

PubMed1985

Inability to detect a K cell in bovine peripheral blood leukocytes.

Antibody-dependent cellular cytotoxicity to viral-infected cells, chicken red blood cells, and tumor cells was tested using different effector cell populations: polymorphonuclear cells, mononuclear cells, and mononuclear cells separated into adherent and nonadherent populations by Sephadex G-10. Polymorphonuclear cells were the most efficient mediators of antibody-dependent cellular cytotoxicity against most targets, although a combination of G-10 adherent and polymorphonuclear cells was more efficient in killing infectious bovine rhinotracheitis virus-infected cells than either single cell population. Removal of G-10 adherent cells from the mononuclear cell population removed all antibody-dependent cellular cytotoxicity from that population, indicating the lack of a typical K cell in bovine peripheral blood.

Animals↗

PubMed1985

Cell-mediated cytotoxicity of bovine mononuclear cells to IBRV-infected cells: dependence on Sephadex G-10 adherent cells.

Following intranasal inoculation of cattle with infectious bovine rhinotracheitis virus (IBRV) mononuclear cells that produced a genetically unrestricted cytotoxic response against IBRV-infected, but not against uninfected cells, were present in peripheral blood. Cytotoxicity was detected between 6 and 14 days after primary infection in a 20 h, but not in a 5 h, 51Cr-release assay. Cytotoxic activity was present in peripheral blood mononuclear cells from infected and subsequently hyperimmunized cattle for a considerably longer time. Neither natural cytotoxicity, antibody-dependent cell cytotoxicity, nor antibody produced during the assay was responsible for the cytotoxicity. However, cytotoxicity was dependent upon an adherent mononuclear cell that was partially removed by passage over nylon wool and completely removed by passage over Sephadex G-10.

Animals↗

PubMed1985

Effect of human leukocyte A interferon on prevention of infectious bovine rhinotracheitis virus infection of cattle.

Human leukocyte A interferon was evaluated for its ability to prevent infectious bovine rhinotracheitis virus-induced respiratory tract disease in cattle. Weanling calves were treated daily for 1 week with 50 X 10(6) U of interferon, intranasally (by nebulization) and IM, and inoculated with infectious bovine rhinotracheitis virus on the first day of treatment. Respiratory tract disease was less severe in treated as compared with nontreated calves which were given only infectious bovine rhinotracheitis virus, and infection in the treated calves occurred later than in the untreated calves. Viral shedding and appearance of viral neutralizing antibodies occurred later in treated calves than in calves given only virus. Because several calves in a treatment group were housed together, whether the late appearance of infection in some interferon-treated calves was due to emergence of suppressed virus or to horizontal transmission from calves shedding virus could not be determined. One calf in the interferon-treated group developed antibody to human interferon and a few treated calves had transient elevation of hepatic enzymes. Interferon-treated calves developed a high temperature which subsided on termination of treatment. Production of disease was considerably dependent on the amount of virus and interferon given, since calves given 300 times more virus and approximately half as much interferon showed no evidence of protection against infection.

Administration, Intranasal↗

PubMed1983

Characteristics of the polyriboinosinic acid:polyribocytidylic acid assay for noncytopathogenic bovine viral diarrhea virus.

The use of polyriboinosinic acid:polyribocytidylic acid (poly I:C) for noncytopathogenic bovine viral diarrhea virus (NC-BVDV) assay (PINBA) was studied. Several viruses were tested for their suitability as test challenge viruses. In addition to vesicular stomatitis virus, which previously was shown to be a suitable challenge virus, bovine enteroviruses also were found to be suitable, whereas infectious bovine rhinotracheitis virus and parainfluenza type 3 virus were only marginally suitable. Bovine embryonic skin (BES) cultures developed resistance to PINBA within a few in vitro passages, whereas bovine embryonic lung (BEL) cultures did not. At certain passages, BES cultures were 500,000 times more resistant than BEL cultures. To determine whether the difference in viral titers on BEL and BES cultures was due to NC-BVDV replication, PINBA and fluorescent antibody assays were compared on the cultures. Resistance of BES cultures to PINBA was not due to an inability of the virus to replicate in the cultures, but was due to an inability of PINBA to detect the virus. Viral titers were comparable by fluorescent antibody assay titers on BES and BEL cultures, but were considerably higher than viral titers on BES cultures with PINBA. Variations in viral titers, using PINBA on BEL cultures, were observed and were considered to be due to cultural conditions, such as the presence of low levels of BVDV antibodies in bovine fetal serum used in the medium. Treatment of BEL cultures with poly I:C or interferon showed that NC-BVDV was sensitive to interferon as determined by virus-yield reduction.

Animals↗

PubMed1982

A microtiter test for detecting and titrating noncytopathogenic bovine viral diarrhea virus.

Bovine cells free of noncytopathogenic bovine viral diarrhea virus (NC-BVDV) treated with polyriboinosinic acid : polyribocytidylic acid (poly I:C) were protected against challenge with vesicular stomatitis virus (VSV), whereas NC-BVDV-infected cells treated with poly I:C were not protected against VSV. An assay based on the ability of NC-BVDV to inhibit poly I:C protection of cells against VSV was developed and is herein referred to as PINBA (poly I:C for NC-BVDV assay). Noncytopathogenic BVDV was titrated as cytopathogenic strains except that several days after infection with NC-BVDV, the cultures were treated with poly I:C and VSV. Titration endpoints were reached 24 hours later. PINBA was standardized for amount of VSV, time of addition of poly I:C, and time NC-BVDV had to be present to obtain stable titration endpoints. PINBA also was useful for titrating virus neutralizing antibodies. Compared with the fluorescent antibody test, PINBA was less subjective for detection of NC-BVDV. Compared with the interference test in which NC-BVDV infected cultures are challenged with a cytopathogenic strain of BVDV, PINBA was more reliable. The technique described herein is a simple and practical microtiter method for titrating NC-BVDV and virus neutralizing antibodies and for the presumptive detection of NC-BVDV.

Animals↗