The "Levy" manuscript was published 12/96, and Galton writes the letter 12/97? After being beaten over the head with it for a year, it looks like perhaps BlueStone got him to do something.
J Immunol 1996 Dec 15;157(12):5503-5511
A nine-amino acid peptide from IL-1beta augments antitumor immune
responses induced by protein and DNA vaccines.
Hakim I, Levy S, Levy R
The idiotypic determinants of B cell lymphoma provide a tumor-specific
Ag and a target for immunotherapy. We have
developed several generations of idiotype vaccines that were tested in
an animal model, the 38C13 mouse B cell lymphoma.
Initially we showed that effective tumor immunity was elicited by the
syngeneic Id when it was conjugated to a carrier protein
and mixed with an adjuvant. A subsequent generation of Id vaccines
eliminated the need for a carrier protein and for an
adjuvant by incorporating cytokines into fusion proteins containing the
Id. A third generation of vaccines consisting of naked
DNA encoding the Id-granulocyte-macrophage colony-stimulating factor
(GM-CSF) fusion proteins was equally effective in
inducing tumor immunity. To determine whether Ig variable regions, in
the absence of constant regions, could be
immunotherapeutic in this model, we tested the use of single-chain Fv
(scFv). scFv proteins, produced in bacteria, and naked
DNA encoding scFv were used in this study. scFv was tested alone or
fused to GM-CSF or an immunoenhancing peptide
derived from IL-1beta. Here we demonstrate that scFv-GM-CSF was
effective only when injected as a protein, not as a DNA
vaccine. In contrast, both scFv-IL-1beta peptide fusion protein and
naked DNA encoding it induced tumor immunity that
protected mice from tumor challenge.
UI - 98031284
AU - Maecker HT
AU - Umetsu DT
AU - DeKruyff RH
AU - Levy S
TI - DNA vaccination with cytokine fusion constructs biases the immune
response to ovalbumin.
AB - DNA vaccination may work through direct transfection of antigen
presenting cells (APC), or by secretion of the encoded protein by
muscle or skin cells for uptake by APC. If cytokines are attached
to
the antigen, they may influence APC or responding T cells to drive
the
response toward a Th1 or Th2 direction, and/or potentiate it in an
antigen-specific manner. To test this concept, expression vectors
were
constructed containing the ovalbumin (OVA) gene either alone, or
linked
to cytokine genes including GM-CSF, IFN-gamma, IL-2, IL-4, IL-12,
or a
sequence encoding nine amino acids of IL-1 beta. These constructs
expressed OVA-cytokine fusion proteins in vitro which retained
cytokine
bioactivity. C57BL/6 mice were injected intramuscularly with the
DNA
constructs. Little if any OVA-specific antibody was produced in
response to any of the DNA constructs, except for OVA-IL-4.
However,
lymphocytes from BALB/c mice vaccinated with OVA-IL-12 and
OVA-IL-1
beta constructs produced more IFN-gamma and less IL-4 during in
vitro
restimulation assays than did other groups. All constructs
elicited OVA-
specific cytotoxic responses which were maintained or even
increased
over 16 weeks. The OVA-IL-12 and OVA-IL-1 beta peptide constructs
elicited the strongest cytotoxic responses at 2 weeks
postinjection.
Cytotoxic responses were seen in all animals, even those lacking
OVA-
specific Ab, and were not related to Ab level. These studies
indicate
that the humoral, cytokine, and cytotoxic responses to DNA
vaccination
can be effectively altered by certain cytokine fusion constructs.
AD - Department of Medicine/Oncology, Stanford University Medical
Center, CA
94305, USA.
SO - Vaccine 1997 Oct;15(15):1687-96 |
