Previous studies have shown that the anionic alkali mineral complex BARODON has an immunoenhancin... more Previous studies have shown that the anionic alkali mineral complex BARODON has an immunoenhancing effect on pigs as an adjuvant and as a nonspecific immunostimulant. Likewise, the equine immune system has been defined with various monoclonal antibodies specific to equine leukocyte differentiation antigens to determine the possibility of enhancing equine resistance to respiratory diseases and promoting other immunostimulatory effects with the application of BARODON. Compared with the control group, after 3 weeks of treatment, BARODON-treated groups showed higher proportions of cells ( P < 0.05) expressing major histocompatibility complex class II and CD2, CD4 + , CD4 + CD25 + , CD8 + , and CD8 + CD25 + T lymphocytes, dendritic cells, and surface immunoglobulin M + B lymphocytes in peripheral blood, as well as enhanced cell proliferative responses with phytohemagglutinin and increased phagocytic activity against Streptococcus equi and Staphylococcus aureus strains with high antibi...
Since the discovery that Mycobacterium aviumsubsp. paratuberculosis(Map) is the causative agent o... more Since the discovery that Mycobacterium aviumsubsp. paratuberculosis(Map) is the causative agent of Johne’s disease (JD) in cattle at the end of the nineteenth century, movement of livestock latently infected with Map has led to the spread of JD throughout the world. A new form of enteritis with clinical features of JD in cattle appeared in humans concurrent with the appearance of Map as a disease problem in livestock. The demonstration that Map is a zoonotic pathogen and the causative agent of the new form of enteritis in humans, however, wasn’t recognized until late in the twentieth century when methods were developed to detect the presence Map in tissues from patients with the new form of clinical enteritis. The objective of this short review is to provide a brief history explaining how Map has become a major disease problem in livestock and humans and then provide a review of the progress that has been made in treating patients with an enteritis caused by Map and the strategies underway to develop a vaccine to control infection in livestock.
In this paper, we describe a technique for using the statistical method of fractional factorial d... more In this paper, we describe a technique for using the statistical method of fractional factorial design in the optimization of an ELISA. Fractional factorial design dramatically reduces the total number of experiments required in the optimization. In addition, this technique enables us to determine the parameters that give the maximum sensitivity range with the most accuracy for the ELISA studied.
Circulating tumor cells (CTCs) in the peripheral blood are shed from either the primary tumor or ... more Circulating tumor cells (CTCs) in the peripheral blood are shed from either the primary tumor or its metastases. These disseminated CTCs are directly responsible for the most cancer deaths and the major cause of prostate cancer-related mortality. Therefore, the earlier detection of CTCs and the ability to remove CTCs from circulation could result in more positive patient outcomes. However, capturing low abundant CTCs from peripheral blood remains a challenge. The cell-surface enzyme prostate-specific membrane antigen (PSMA) is an important biomarker in prostate cancer research. Its upregulation on prostate tumor cells has attracted significant attention as a target for imaging and therapeutic agents. We recently developed a technology to enrich CTCs from a large blood volume by combining apheresis and elutriation technologies with a CTC selection technology based on immobilized high-affinity irreversible PSMA inhibitor. The commercial centrifugal apheresis and elutriation processes can concentrate PCTCs from large blood volumes in a standard elutriator. In this presentation, we will describe a selective and sensitive methodology to enrich, capture, and quantify metastatic prostate cancer cells. Citation Format: Lisa Y. Wu, Dana E. Abernathy, TianCheng Liu, Bernard J. Van Wie, William C. Davis, Clifford E. Berkman. Large-scale isolation of prostate cancer cells by combining leukapheresis and elutriation with chemoaffinity capture [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr C40.
Studies with Mycobacterium avium subsp. paratuberculosis (Map) in cattle revealed deletion of rel... more Studies with Mycobacterium avium subsp. paratuberculosis (Map) in cattle revealed deletion of relA, a global regulator gene, abrogated ability of the mutant to establish a persistent infection, attributed to development of an immune response that cleared infection. Analysis of the recall response demonstrated presence of CD8 cytotoxic T cells that kill intracellular bacteria. Replication of the primary response demonstrated the CTL response could be elicited with the ΔMap/relA mutant or the target of the immune response, a 35 kD membrane protein. Follow up comparative studies with Mycobacterium bovis bacillus Calmette-Guérin (BCG) and a BCG relA (ΔBCG/relA) deletion mutant revealed deletion of relA enhanced the CTL response compared to BCG. Analysis of the cytokine profile of cells proliferating in response to stimulation with BCG or BCG/relA showed increased expression of IFN-γ, TNF-α, and IL-17 by cells stimulated with ΔBCG/relA in comparison with BCG. The proliferative and CTL responses were markedly reduced in response to stimulation with heat killed BCG or ΔBCG/relA. Intracellular bacterial killing was mediated through the perforin, granzyme B (GnzB), and the granulysin pathway. The data indicate relA is the Achilles' heel for pathogenic mycobacteria and deletion may be key to improving efficacy of attenuated vaccines for mycobacterial pathogens.
Control of Johne's disease, caused by Mycobacterium avium subsp. paratuberculosis, has been d... more Control of Johne's disease, caused by Mycobacterium avium subsp. paratuberculosis, has been difficult because of a lack of an effective vaccine. To address this problem we used targeted gene disruption to develop candidate mutants with impaired capacity to survive ex vivo and in vivo to test as a vaccine. We selected relA and pknG, genes known to be important virulence factors in Mycobacterium tuberculosis and Mycobacterium bovis, for initial studies. Deletion mutants were made in a wild type Map (K10) and its recombinant strain expressing the green fluorescent protein (K10-GFP). Comparison of survival in an ex vivo assay revealed deletion of either gene attenuated survival in monocyte-derived macrophages compared to survival of wild-type K10. In contrast, study in calves revealed survival in vivo was mainly affected by deletion of relA. Bacteria were detected in tissues from wild-type and the pknG mutant infected calves by bacterial culture and PCR at three months post infectio...
Previous studies have shown that the anionic alkali mineral complex BARODON has an immunoenhancin... more Previous studies have shown that the anionic alkali mineral complex BARODON has an immunoenhancing effect on pigs as an adjuvant and as a nonspecific immunostimulant. Likewise, the equine immune system has been defined with various monoclonal antibodies specific to equine leukocyte differentiation antigens to determine the possibility of enhancing equine resistance to respiratory diseases and promoting other immunostimulatory effects with the application of BARODON. Compared with the control group, after 3 weeks of treatment, BARODON-treated groups showed higher proportions of cells ( P < 0.05) expressing major histocompatibility complex class II and CD2, CD4 + , CD4 + CD25 + , CD8 + , and CD8 + CD25 + T lymphocytes, dendritic cells, and surface immunoglobulin M + B lymphocytes in peripheral blood, as well as enhanced cell proliferative responses with phytohemagglutinin and increased phagocytic activity against Streptococcus equi and Staphylococcus aureus strains with high antibi...
Since the discovery that Mycobacterium aviumsubsp. paratuberculosis(Map) is the causative agent o... more Since the discovery that Mycobacterium aviumsubsp. paratuberculosis(Map) is the causative agent of Johne’s disease (JD) in cattle at the end of the nineteenth century, movement of livestock latently infected with Map has led to the spread of JD throughout the world. A new form of enteritis with clinical features of JD in cattle appeared in humans concurrent with the appearance of Map as a disease problem in livestock. The demonstration that Map is a zoonotic pathogen and the causative agent of the new form of enteritis in humans, however, wasn’t recognized until late in the twentieth century when methods were developed to detect the presence Map in tissues from patients with the new form of clinical enteritis. The objective of this short review is to provide a brief history explaining how Map has become a major disease problem in livestock and humans and then provide a review of the progress that has been made in treating patients with an enteritis caused by Map and the strategies underway to develop a vaccine to control infection in livestock.
In this paper, we describe a technique for using the statistical method of fractional factorial d... more In this paper, we describe a technique for using the statistical method of fractional factorial design in the optimization of an ELISA. Fractional factorial design dramatically reduces the total number of experiments required in the optimization. In addition, this technique enables us to determine the parameters that give the maximum sensitivity range with the most accuracy for the ELISA studied.
Circulating tumor cells (CTCs) in the peripheral blood are shed from either the primary tumor or ... more Circulating tumor cells (CTCs) in the peripheral blood are shed from either the primary tumor or its metastases. These disseminated CTCs are directly responsible for the most cancer deaths and the major cause of prostate cancer-related mortality. Therefore, the earlier detection of CTCs and the ability to remove CTCs from circulation could result in more positive patient outcomes. However, capturing low abundant CTCs from peripheral blood remains a challenge. The cell-surface enzyme prostate-specific membrane antigen (PSMA) is an important biomarker in prostate cancer research. Its upregulation on prostate tumor cells has attracted significant attention as a target for imaging and therapeutic agents. We recently developed a technology to enrich CTCs from a large blood volume by combining apheresis and elutriation technologies with a CTC selection technology based on immobilized high-affinity irreversible PSMA inhibitor. The commercial centrifugal apheresis and elutriation processes can concentrate PCTCs from large blood volumes in a standard elutriator. In this presentation, we will describe a selective and sensitive methodology to enrich, capture, and quantify metastatic prostate cancer cells. Citation Format: Lisa Y. Wu, Dana E. Abernathy, TianCheng Liu, Bernard J. Van Wie, William C. Davis, Clifford E. Berkman. Large-scale isolation of prostate cancer cells by combining leukapheresis and elutriation with chemoaffinity capture [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr C40.
Studies with Mycobacterium avium subsp. paratuberculosis (Map) in cattle revealed deletion of rel... more Studies with Mycobacterium avium subsp. paratuberculosis (Map) in cattle revealed deletion of relA, a global regulator gene, abrogated ability of the mutant to establish a persistent infection, attributed to development of an immune response that cleared infection. Analysis of the recall response demonstrated presence of CD8 cytotoxic T cells that kill intracellular bacteria. Replication of the primary response demonstrated the CTL response could be elicited with the ΔMap/relA mutant or the target of the immune response, a 35 kD membrane protein. Follow up comparative studies with Mycobacterium bovis bacillus Calmette-Guérin (BCG) and a BCG relA (ΔBCG/relA) deletion mutant revealed deletion of relA enhanced the CTL response compared to BCG. Analysis of the cytokine profile of cells proliferating in response to stimulation with BCG or BCG/relA showed increased expression of IFN-γ, TNF-α, and IL-17 by cells stimulated with ΔBCG/relA in comparison with BCG. The proliferative and CTL responses were markedly reduced in response to stimulation with heat killed BCG or ΔBCG/relA. Intracellular bacterial killing was mediated through the perforin, granzyme B (GnzB), and the granulysin pathway. The data indicate relA is the Achilles' heel for pathogenic mycobacteria and deletion may be key to improving efficacy of attenuated vaccines for mycobacterial pathogens.
Control of Johne's disease, caused by Mycobacterium avium subsp. paratuberculosis, has been d... more Control of Johne's disease, caused by Mycobacterium avium subsp. paratuberculosis, has been difficult because of a lack of an effective vaccine. To address this problem we used targeted gene disruption to develop candidate mutants with impaired capacity to survive ex vivo and in vivo to test as a vaccine. We selected relA and pknG, genes known to be important virulence factors in Mycobacterium tuberculosis and Mycobacterium bovis, for initial studies. Deletion mutants were made in a wild type Map (K10) and its recombinant strain expressing the green fluorescent protein (K10-GFP). Comparison of survival in an ex vivo assay revealed deletion of either gene attenuated survival in monocyte-derived macrophages compared to survival of wild-type K10. In contrast, study in calves revealed survival in vivo was mainly affected by deletion of relA. Bacteria were detected in tissues from wild-type and the pknG mutant infected calves by bacterial culture and PCR at three months post infectio...
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