>> Do yourself a favor, stick to what you know buddy, and you will save yourself some money (and anguish) <<
OK......
Proc Natl Acad Sci U S A 1994 Aug 2;91(16):7727-31
A mutant epidermal growth factor receptor common in human glioma
confers enhanced tumorigenicity.
Nishikawa R, Ji XD, Harmon RC, Lazar CS, Gill GN, Cavenee WK, Huang HJ
Ludwig Institute for Cancer Research, La Jolla, CA 92093-0660.
The development and neoplastic progression of human astrocytic tumors appears to result through an accumulation of genetic
alterations occurring in a relatively defined order. One such alteration is amplification of the epidermal growth factor receptor
(EGFR) gene. This episomal amplification occurs in 40-50% of glioblastomas, which also normally express endogenous receptors.
Moreover, a significant fraction of amplified genes are rearranged to specifically eliminate a DNA fragment containing exons 2-7
of the gene, resulting in an in-frame deletion of 801 bp of the coding sequence of the extracellular domain. Here we used retroviral
transfer of such a mutant receptor (de 2-7 EGFR) into glioblastoma cells expressing normal endogenous receptors to test whether
the mutant receptor was able to augment their growth and malignancy. Western blotting analysis showed that these cells
expressed endogenous EGFR of 170 kDa as well as the exogenous de 2-7 EGFR of 140-155 kDa. Although holo-EGFRs were
phosphorylated on tyrosine residues only after exposure of the cells to ligand, de 2-7 EGFRs were constitutively phosphorylated. In
tissue culture neither addition of EGF nor expression of the mutant EGFR affected the rate of cell growth. However, when cells
expressing mutant EGFR were implanted into nude mice subcutaneously or intracerebrally, tumorigenic capacity was greatly
enhanced. These results suggest that a tumor-specific alteration of the EGFR plays a significant role in tumor progression perhaps
by influencing interactions of tumor cells with their microenvironment in ways not easily assayed in vitro.
Am J Pathol 1994 Jul;145(1):33-6
Anti-CD31 delays platelet adhesion/aggregation at sites of endothelial injury
in mouse cerebral arterioles.
Rosenblum WI, Murata S, Nelson GH, Werner PK, Ranken R, Harmon RC
Department of Pathology (Neuropathology), Virginia Commonwealth University, Medical College of Virginia, Richmond
23298-0017.
The arterioles on the surface of the mouse brain (pial arterioles) were observed by in vivo microscopy. A focus of minor
endothelial damage was produced in a single pial arteriole in each mouse by briefly exposing the site to a helium neon laser after
an intravenous injection of Evans blue. Mice were injected 10 minutes before injury with a monoclonal antibody (MAb) to mouse
CD31, also known as platelet endothelial cell adhesion molecule. This treatment doubled (P < .01) the time required for the laser to
produce a recognizable platelet aggregate. In additional experiments, an MAb to mouse CD61 and an MAb to mouse intercellular
adhesion molecule 1 had no effect. The data support previous observations indicating that platelet adhesion/aggregation in this
model is induced by endothelial injury without exposure of basal lamina. The data are consistent with the hypothesis that the
endothelial injury exposes or activates a platelet endothelial cell adhesion molecule on the endothelium which is blocked by the
MAb directed against CD31. This may be the first demonstration of an effect of an anti-platelet endothelial cell adhesion molecule
on platelet endothelial cell adhesion molecule on platelet adhesion/aggregation in vivo.
Infect Immun 1989 Jul;57(7):1936-41
Monoclonal antibody-mediated protection and neutralization of motility in
experimental Proteus mirabilis infection.
Harmon RC, Rutherford RL, Wu HM, Collins MS
Research and Development, Cutter-Biological, Berkeley, California.
A panel of monoclonal antibodies with specificity for a wound isolate of Proteus mirabilis was established. Of nine antibodies
studied in detail, three were broadly reactive with various Proteus isolates, while six reacted in a serotype-specific fashion with the
strain used for immunization. Five of the six serotype-specific antibodies were reactive with lipopolysaccharide. The sixth
serotype-specific antibody, 4-F (immunoglobulin G1 [IgG1]), was potently protective in a burn wound sepsis model and recognized
a protein antigen. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot (immunoblot) analysis were used to
determine that 4-F was reactive with flagellar protein. Approximately 1.3 micrograms of the antibody was sufficient to provide
protection against 8 50% lethal doses of wound isolate, and approximately 26 micrograms provided full protection against challenge
with 333 50% lethal doses. In vitro test results indicated that 4-F inhibited the motility of the wound isolate, and in vivo testing
showed that it inhibited dissemination of the inoculum from the burn site to the liver and spleen. Whereas the antibody was highly
effective in preventing the death of mice subsequent to challenge at a burn site, no protection was seen following an intraperitoneal
challenge. These results may therefore indicate that the protection observed in the burn model is solely a reflection of the capacity
of 4-F to neutralize bacterial motility.
Immunogenetics 1986;23(6):406-8
A new murine lymphocyte alloantigen, Ly-27.2.
Matsushima GK, Harmon RC, Frelinger JA
Vox Sang 1984;47(6):412-20
Detection of IgG-associated determinants in reduced and alkylated
preparations of human IgG3 by monoclonal antibodies.
Brown AM, Dumas ML, Reimer CB, Louie RE, Harmon RC
Using classical typing antisera, previous experiments have failed to demonstrate IgG3 in partially reduced and alkylated
preparations of human IgG intended for intravenous application (IGIV). To establish that IgG3 is actually present in such
preparations, we designed an enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies as solid-phase reagents
and protein A-purified IgG3 as antigen. Three different samples of reduced and alkylated antigen were used: (1) IgG3 isolated
from a ready-for-infusion IGIV; (2) IgG3 which was purified from an intramuscular (Cohn fraction II) IgG solution before being
subjected to a mild reduction and alkylation procedure, and (3) completely reduced and alkylated IgG3. The reduction and
alkylation procedure did not affect the solubility of IgG3, indicating that IGIV prepared in this manner should contain normal
quantities of IgG3. In the ELISA, solid-phase monoclonals which were cross-reactive with multiple IgG subclasses clearly reacted
with reduced and alkylated IgG3. Furthermore, there was no substantial difference between the quantities of modified and native
antigen required for 50% maximal ELISA signal. In contrast, solid-phase monoclonals with IgG3-restricted specificity did not
recognize reduced and alkylated material. These results indicate that IGIV prepared by reduction and alkylation has a normal IgG3
content and confirm that some IgG3-specific determinants are altered by the modification procedure.
Virology 1983 Dec;131(2):296-307
Antigenic relationships of murine coronaviruses: analysis using monoclonal
antibodies to JHM (MHV-4) virus.
Fleming JO, Stohlman SA, Harmon RC, Lai MM, Frelinger JA, Weiner LP
Monoclonal antibodies were produced to JHMV-DL, a neurotropic member of the mouse hepatitis virus (MHV) or murine
coronavirus group. Of 23 antibodies isolated, 10 were specific for the major envelope glycoprotein, gp180/90, 10 for the
nucleocapsid protein, pp60, and 3 for the minor envelope glycoprotein, gp25. Eleven different MHV isolates were used in antibody
binding assays to study antigenic relationships among the viruses. Each MHV isolate tested had a unique pattern of antibody
binding, indicating that each is a distinct strain. Conservation of JHMV-DL antigenic determinants varied among the three proteins,
with pp60 showing intermediate conservation, gp180/90 little conservation, and gp25 marked conservation in the different MHV
strains. Monoclonal antibodies to pp60 proved most useful in delineating antigenic relationships among MHV strains. These
antigenic groups correlated with pathogenic types, indicating that pp60 may be one of the gene products which mediates the
distinct disease patterns manifested by different murine coronaviruses.
Int J Cancer 1983 Mar 15;31(3):309-14
Natural killer cell activity during mouse hepatitis virus infection: response in
the absence of interferon.
Stohlman SA, Brayton PR, Harmon RC, Stevenson D, Ganges RG, Matsushima GK
The ability of the JHM3 strain of mouse hepatitis virus (MHV) to induce natural killer (NK) cells was examined. Infection of
C57BL/6 (B6) mice with this virus resulted in the augmentation of natural cytotoxicity against YAC-I target cells in the absence of
a detectable interferon response. The cells responsible for this increased cytotoxicity were sensitive to complement-mediated lysis
with an anti-Q-5 reagent but not with a Thy 1.2 antiserum, indicating that they possess an NK-like surface phenotype. Although
variation in the NK response of individual B6 mice following JHM virus infection was found, even the animal with the most
responsive NK cell population had no detectable interferon in the spleen. This finding contrasted with observations with an
unrelated virus (lymphocytic choriomeningitis virus) and a serologically related strain of MHV. Infection with both of these viruses
induced augmented NK cell activity and interferon responses. In addition, we found that neither the ability to mount an augmented
NK cell response nor preferential lysis of virus-infected targets correlated with resistance or susceptibility to JHM virus infection.
Immunogenetics 1983;18(5):541-5
Monoclonal antibodies reactive with H-2 determinants.
Harmon RC, Stein N, Frelinger JA
Nature 1982 Jun 3;297(5865):415-7
Product of a transferred H-2Ld gene acts as restriction element for
LCMV-specific killer T cells.
Orn A, Goodenow RS, Hood L, Brayton PR, Woodward JG, Harmon RC, Frelinger JA
Proc Natl Acad Sci U S A 1982 Jun;79(11):3613-7
Specific recognition of the product of a transferred major histocompatibility
complex gene by cytotoxic T lymphocytes.
Woodward JG, Orn A, Harmon RC, Goodenow RS, Hood L, Frelinger JA
Mouse L cells transfected with a genomic clone containing the H-2Ld gene (8-5 cells) were shown to function as targets for
H-2Ld-specific cytotoxic T lymphocytes (CTL). The CTL-mediated lysis of 8-5 cells was shown to be H-2Ld specific by the use
of (i) CTL with restricted reactivity, (ii) unlabeled target inhibiton, and (iii) monoclonal antibody inhibiton. We also demonstrated
that 8-5 cells could function as targets for antibody-plus-complement-mediated cell lysis. Specificity was confirmed by using
H-2Ld-specific monoclonal antibodies. These experiments demonstrate that the gene products of a major histocompatibility
complex genomic clone can be functionally expressed in a foreign cell and can mediate immunologically specific cellular
interactions.
(getting tired, so I'll stop there)
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