STANFORD UNIVERSITY
STANFORD, CALIFORNIA

DEPARTMENT OF MEDICAL MICROBIOLOGY November 11 > 1958

Dr. Joshua Lederberg
Genetics Building
University of Wisconsin
Madison 6, Wisconsin

Dear Joshua:

I am amazed that, at a time like this when ex-
citement must be rampant without and within, you can take
time to write a thoughtful letter about the hypersensitivity
questions in which you are interested. Your questions are
good ones, and I will try to do justice to them.

I sent you the manuscript because I believe that
it may pertain to the question of earliest cell response to
immunologic stimuli which interests you. We made a special
report of this work, not because the findings are original
in the sense of a new discovery, but in order to try to put
into perspective what these reactivities consist of. Uhr
and Pappenheimer have been speaking of the induction of
delayed hypersensitivity by means of antigen-antibody
complexes, as if such complexes in the region of antibody
excess have some special property in this respect. Our
work was simply to demonstrate, as has been done less
pointedly by others in the past, that the reactivity which
they see is one which is occasioned by an antigenic stimulus
alone; if this is small, the evanescent delayed-appearing
response appears to be the only one which the animal is
capable of. If it is larger, this is succeeded by humoral
antibodies and Arthus (immediate) reactivity.

You ask whether this work is related to that
of Tremaine. Her work suggests that animals which have
been immunized with protein in a manner calculated to pro-
duce circulating antibodies and immediate hypersensitivity
can transfer, by means of peritoneal exudate cells, delayed
reactivity to normal recipient rabbits. If this is true,
it would mean that all animals immunized with ordinary
protein antigens actually have a degree of delayed hyper-
sensitivity at all times, and that this does not show up
Dr. Lederberg 2 November 11, 1958

perhaps when the immunized animal itself is skin-tested
later in its course because the larger immediate reactions
obscure the delayed ones. Early in the game, before anti-
bodies and immediate reactivity have appeared, the delayed
reactions can be seen. If all this is true, it would mean
that what we call the Jones-Mote type of early appearing
evanescent delayed hypersensitivity actually runs through-
out the period of immunologic response and in the sense

of persistence would probably be the same as conventional
delayed hypersensitivity. Whether this is true or not,
for purposes of your interest in the matter it still re-
mains rather apparent that the delayed hypersensitive kind
of reactivity seems to be the first response of the animal
to an antigen. But to take the matter further, I am not
convinced as yet that the "Jones-Mote" delayed hypersensi-
tivity runs throughout the period of immunologic response
as Tremaine's work suggests. If this were true, one should
expect to be able to show corneal reactions (which everyone
agrees occur only when delayed hypersensitivity is present)
in any animal which has been immunized with any antigen,
and this is not so, as evidenced by the fact that the cor-
neal test is a very good one used to distinguish immediate
from delayed hypersensitive states. In our own work with
guinea pigs highly anaphylactic to tuberculoprotein (when
lipopolysaccharide was not used with it) no corneal res-
ponses were seen at all, and this has been general experi-
ence. My point is that if she can transfer cells to a
recipient animal intravenously and by this means make the
cornea of the recipient animal reactive to antigen, why
can't the donor of the cells itself show the same kind of
reactivity? There is other evidence, but I don't want to
prolong this letter to the length of a manuscript.

You ask also how all this--Tremaine's and our
own work--may be related to the observations of Rosenberg
et al. in a recent issue of the Journal of Immunology. In
that case cells of the spleen were transferred to skin
sites and later injections of antigen were made into the
same sites for the purpose of demonstrating the appearance
of small amounts of antibodies manufactured by the trans-
planted cells. The essential conditions here are the same
Dr. Lederberg 3 November 11, 1958

as those used by Tremaine, and this confuses the issue. I
should say, however, that Rosenberg was observing true im-
mediate reactions for the simple reason that they occurred
immediately. The animals were killed fifteen minutes after
antigen injection in order to determine the extent of in-
filtration of the injected site by dye. If Tremaine's work
holds up and applies to the skin as well, which it should,
I would say that if Rosenberg were to wait for an appropri-
ate period of hours after the injection of test antigen

he might see the same kind of reactions that Tremaine saw
in the corneas. I might add also that the small numbers

of cells used by Rosenberg et al. apparently cannot trans-
fer delayed hypersensitivity. At least all instances
described make use of the cells pooled from several donors
to sensitize one recipient, whereas Rosenberg used cells
from one recipient to sensitize a number of donor sites.

So far I have only replied to your first para-
graph. Your next one takes up the question as to whether
perhaps only a single molecular kind of antibody is pro-
duced no matter what the inciting conditions (at least
that is how I interpret your statement), but that this
antibody must be closely regulated in amount and timing
of release if it is to be totally absorbed by the tissues
leaving no detectable amounts in the serum. I don't think
that this is so. If one uses, let us say, a protein with
lipopolysaccharide good antibody induction as well as de-
layed hypersensitivity apparently to the same antigen is
produced almost simultaneously. Of course, it is true
that these proteins are not single molecular species and
one could say that the delayed response is directed against
perhaps a minor component of the tuberculoprotein whereas
the antibodies are directed against some other component
or components, but this doesn't seem likely.

You next take up the matter of Brent's work.
Our paper is not intended to criticize the viewpoint that
the homograft immunity may be a form of delayed hypersen-
sitivity. Brent may well be right; a recent paper by him
and others in the Lancet, September 13, 1958, page 561,
presents pretty good evidence that perhaps the reactivity
Dr. Lederberg 4 November 11, 1958

is of this type in guinea pigs. These kinds of observations,
of course, fail to account for the observed fact that
agammaglobulinemic patients, who are virtually unable to
produce antibodies, but can develop delayed hypersensitivity,
still are able to accept homografts, a fact which would

make it appear that antibodies are important in the anti-
homograft reaction, at least in the human being. I suppose
that the solution to this problem has not yet been found.

I believe that it is perfectly feasible that
the H, genes may reflect different antigenic substances
and that tissues other than erythrocytes may possess an
antigen or antigens which the erythrocytes don't have.
There are many examples of this; a well-known one is the
case of the Forssman antigen in guinea pigs. This occurs
in all tissues except the red blood cells. But I think
that your idea that Brent should try injections of erythro-
cytes along with a heterologous tissue in order to find
whether a T-provoking "principle" might be present in
such tissue is a good one. He has tried red cells along
with lipopolysaccharide and tubercle bacilli; he was up
to see me a couple of years ago while working with Owen
at Cal Tech, and obtained material for this purpose. The
results were negative.

I believe that Brent and his co-workers have
tried to "unify" their work with other work in immunology
and allergy. The trouble is that unification has so far
been very difficult, for others in addition to Brent. For
example, many people have had a good deal of difficulty in
the past fifty years in trying to determine definitively
whether or not a cellular immunity exists to the tubercle
bacillus, in contradistinction to the antibodies which are
of course found during the course of the infection. The
problem is an analogous one, and people steeped in con-
ventional immunology have so far failed to answer it satis-
factorily.

I too look forward to being able to discuss
these questions with you firsthand. Meanwhile, if you have
Dr. Lederberg 4 November 11, 1958

the time and inclination, I should like very much to see
whether we can communicate effectively on paper.

Best regards to you and your wife.

Sincerely yours,
YY

Ak

Sree ene
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Sidney Raffel

SR: RB

P.S. I just received an extra copy of pageproof; you
needn't bother about the manuscript.

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