The 1ournal of

ALLERGY
and
CLINICAL IMMUNOLOGY
VOLUME 54 NUMBER I

Transfer factor--a theoretical discussion*

F. M. Burner, M.D. P a r k v i U e , V i c t o r i a , A u s t r a l i a

Widespread recent ~ t e r e s t in the transfer factor of H. S. Lawrence and its possible

therapeutic applications has not yet led to an interpretation of the phenomenon
that can bring it into a satisfactory relationship with standard immunologioal theory.
The specifioity of transfer factor ( T F ) has been demonstrated in the sense that
only a small proportion o f the lymphooytes in a recipient can respond to a given
T F preparation. The active agent in T F is known to be o f low molecular weight
(4,000 to 5,000), but its chemical nature and the basis o f its specificity are unknown.
Neither is anything known about the cell receptor that ma~st be capable o f specific
reaction with a partiotlar TF. A n hypothesis is developed which sees the biological
function of T F as to recruit T lymphocytes to deal with situations such as a delayed
hypersensitivity or allograft rejection reaction for which there are inadequate
numbers o f immunologically reactive lymphoeytes. Although the specificity o f T F
does not appear to be directly related to immunological specifioity, it oalls for a
genetically based generation o f diversity roughly analogous to that required in the
develop~nent of the immune system.

T r a n s f e r factor ( T F ) is a phrase introduced by Lawrence 1 in 1955 as a

short name for soluble material he obtained f r o m leukocytes t h a t had been taken
f r o m tuberculin-sensitive donors. W i t h it he could t r a n s f e r skin reactivity of
Mantoux t y p e to volunteer recipients who had previously failed to react to
tuberculin. I t took ten years or more before immunologists took Lawrence's
results s e r i o u s l y - - p e r h a p s largely because they could find no place in the corpus
of theory that was so successfully dealing with the rest of immunological
phenomena. Another reason for skepticism was that what a p p e a r e d to be similar
experiments could not be made to work in guinea pigs or other laboratory

From the School of Microbiology, University of Melbourne, Parkville, Victoria 3052,

Australia.
Received for publication Oct. 19, 1973.
Reprint requests to: F. M. Burnet, M.D., School of Microbiology, University of Melbourne,
Parkville, Victoria 3052, Australia.
*Submitted for publication on the occasion of having received an Honorary Fellowship from
the American Academy of Allergy on Jan. 22, 1974.
Vol. 5d, No. 1, pp. 1-13
2 Burnet J. ALLERGY CLIN. IMMUNOL.
JULY 1974

Special abbreviations used

A_D: antigenic determinant
Ag: antigen
ARC : antigen-reactive cells
CS : combining site
DH : delayed hypersensitivity
DNA: deoxyribonucleic acid
DTF: active dialysate
IR: immune response genes
MIF: migration inhibitory factor
MR : minireceptor
MW: molecular weight
PHA : phytohemagglutinin
PPD: purified protein derivative
RNA : ribonucleic acid
TB : tuberculosis
TF: transfer factor
TR: T cell-immune receptor

animals. However, Lawrence persisted, found good collaborators, and by the

mid-1960's no one doubted the reality of the phenomena. Then, 2 or 3 years ago,
the immunopathologists became interested. Patients with Boeck's sareoid were
ineffective as recipients, usually showing no skin reactions to P P D a f t e r being
given the s t a n d a r d t r a n s f e r factor preparations. Patients with active Hodgkin's
disease were even less responsive. Then. quite recently, but a l r e a d y confirmed,
children with two immunodeficiency diseases, generalized mucocutaneous
candidiasis and the Wiskott-Aldrich syndrome, showing similar a n e r g y to skin
sensitization could regain reactivity and in a proportion of cases show evidence
of clinical i m p r o v e m e n t a f t e r administration of T F . These results have also
ensured t h a t a n y other conditions in which there a p p e a r s to be a failure to
handle a chronic infection because of weakness of T cell function should also
be looked at for possible therapeutic applications of TF.
A t the present time there is a certain skepticism about "cures" of im-
munodeficiency disease, but there is nothing equivocal about the recovery of
skin reactivity to a p p r o p r i a t e antigens a f t e r administration of TF. To me, and
I suspect to most immunologists, the m a j o r challenge of Lawrence's results is to
find a place f o r them within the bounds of orthodox immunological theory. This
p a p e r is essentially an a t t e m p t to construct an hypothesis that, hopefully, could
cover the established facts and be integrated without too much d i ~ c u l t v into
the general body of immunology.

NATURf OF THE PHENOMENON

W o r k on T F is almost wholly concerned with skin sensitizatio~ to nnero-
organismal or chemical antigens, with sensitization to tuberculin as p r o t o t y p e
of the whole series of D H reactors. Other s t a n d a r d antigens used in c|inieal work
are derived f r o m Candi~ albicans, Coceidioides, Histoplasma, the leprosy
bacillus, and m u m p s virus, as well as diphtheria toxoid. Most of Lawrence's
initial work was done with tuberculin sensitization, a n d it is convenient to
VOLUME 54 Transfer factor---a theoretical discussion 3
NUMBER 1

center discussion on this example. DH reactions, as exemplified by the Mantoux

reaction to PPD, are now accepted as being unrelated to the presence of circulat-
ing antibody. In guinea pig experiments, sensitization can be transferred to
another animal only by the use of ly~nphocytes from the activity-sensitized
donor. The lymphocytes concerned in all TF reactions are assumed on rather
uncritical evidence to be thymus-dependent T cells.
One aspect of the DH reaction that is important in the present context is
the development of cytotoxicity that follows activation of lymphocytes in various
ways. This is often, but not always, associated with the liberation of soluble
substances with a variety of names that have toxic or stimulating properties.
These are often referred to generically as lymphokines. A particularly important
type of action is the capacity to stimulate other lymphocytes to blast transforma-
tion, and to the production of similar lymphokines in their turn. The current
interpretation of the DH skin reaction, and of other ways of demonstrating
cell-mediated immunity, is that only a small proportion of specifically reactive
lymphocytes are concerned; 90 per cent or more of the lymphocytes that ac-
cumulate in the region are unsensitized but have been recruited into activity by
secondary results of the specific reactions of a small minority. The details of the
process are far from being fully understood, but its function in recruiting other
cells into useful activity seems to be doubted by nobody. An attempt to interpret
this recruitment is a key feature in what I want to present about TF.
COMPOSITION AND ACTIVITIES OF TF
There have been a number of recent reviews of the general properties of TF, 2"4
and, as this contribution is primarily a theoretical study, the relevant findings
need only be briefly enumerated.
Any extract of mixed human leukocytes will contain a complex mixture of
substances, and even the active dialysate (DTF) from the extract inevitably
contains a wide range of molecular species, including products of enzymatic
breakdown of cellular macromolecules. Peptides and oligonucleotides are present
and it has been claimed that the active agent is a compound containing 12
amino acid residues and 3 nucleotides2 In view of the specificity of the
phenomenon, TF must be present in a number of structurally distinct patterns.
So far, no serious suggestion has been made as to the chemical basis of that
specificity, but, obviously, peptide-nucleotide complexes of MW 4,000-5,000 could
be made up in a large enough range of patterns to fill all requirements. This
may be a convenient mental scaffolding to use in thinking about TF, but not
until two or more distinct specific forms of TF have been chemically defined
will it be legitimate to speak in more than functional terms of the active
molecules concerned.
The degree of specificity has not yet been defined. In general, if a donor is
sensitive to antigens A, B, and C, but not to D, E, and F, a recipient initially
nonreactive to them all will acquire all or some of A, B, C, but never D, E, or F.
At least one exception to the rule has already been reported. Bloom8 describes
one patient who, after receiving TF, developed skin sensitivity to trichophytin,
a reaction shown by none of the donors from whose leukocytes the TF prepara-
tion was made.
4 Burner J. ALLERGY CLIN. IMMUNOL.
JULY 1974

We can reasonably assume that there is only one general type of active
molecule present, necessarily capable of being in a range of specific molecular
patterns in some degree reminiscent of an antibody or a histoeompatibility
antigen. It is essentially an act of faith, supported by an intuitive sense of its
probability, to assume that the phenomena shown by DTF in vitro are due to
the same molecular populations as are responsible for in vivo transfer of D H
and for a therapeutic effect in patients with Wiskott-Aldrich syndrome. There
are two types of in vitro reactions that have been reported. When D T F from
donors reactive to tuberculin is added in vitro to lymphocytes from either
sensitized or normal subjects, no blast transformation takes place. With PPD,
some 2 to 3 per cent of cells in the sensitized category undergo blast transforma-
tion and proliferate: only a trivial response is seen with normal lymphocytes.
With P P D plus DTF, both sensitized and normal lymphocytes show the 2 to 3
per cent of transformed lymphocytesJ The second approach is to demonstrate
the production of MIF or other lymphokines by normal lymphocytes treated
first with TF and then with the specific antigen. In studies of minor immuno-
deficiency states with chronic mucocutaneous candidiasis, several groups 8' 9
have found that recovery of skin reactivity to Candida antigen is associated with
capacity of lymphocytes to respond to antigen with MIF production but not
with lymphoblast transformation. A similar finding m reported by Baram and
Condoulis 1~ in rhesus monkeys sensitized with hemocyanin.
Everything indicates that T F is associated only with activities conventionally
ascribed to lymphocytes of the T system or to cell-mediated immunity. Simply
in confirmation of this it is expedient to record that in congenital sex-linked
agammaglobulinemia, in which there is no evidence of T cell anomalies, there
are equally no abnormal aspects of T F phenomena. T F preparations from
normal PPD-positive individuals confer corresponding skin sensitization on
agammaglobulinemic patients, and vice versa. 2
Among defined immunodeficiencies, the most clear-cut findings come from
investigation of patients with the Wiskott-Aldrich syndrome2.9, 11 The disease
is a genetic one, an X-linked recessive condition affecting only boys and
manifested by thrombocytopenia, eczema, and a high susceptibility to a variety
of infections. Many die early from malignant disease, often of lymphoreticular
type.
Immunologically, the important features concern cell-mediated immunity.
There are no positive skin reactions to the commonly used antigens, and the
children cannot be sensitized with substances like dinitroehlorobenzene. Lympho-
c ~es show normal blast transformation when treated with phytohemagglutinin,
but the response to appropriate antigens is consistently negative or very poor.
MIF is not produced by addition of any of the standard antigens. The main
abnormality reported on the humoral side is a poor or absent production of
antibody against antigens with polysaccharide determinants.
The results of treatment by injections of D T F have shown a good clinical
response in about 50 per cent, with a considerably higher proportion showing
appearance of skin reactivity (13/18). 8 In nearly all instances there was no
response in vitro to stimulation by antigen as judged by incorporation of
VOLUME 54 Transfer factor--a theoretical discussion 5
NUMBER 1

tritiated thymidine, although the MIF lymphokine response usually became

normal. In one well-studied case it could be shown that the capacity to respond
to a given antigen faded within a month or two after the injection of T F and
that episodes of infection were then likely to recur. Both could be corrected by
another injection. Less extensive reports have been published of positive effects
of TF administration in some other immunodeficiency conditions.
There are a number of conditions, probably acquired but perhaps with a
significant genetic component as well, that are well known to show diminished
or absent skin reactions to all the standard reagents. They include sareoidosis, TM
established cancer/s Hodgkin's disease, 14 and gross nutritional conditions such
as kwashiorkor2 s In a table sent to me by Lawrence, the collected results from
studies of such patients given injections either of donor lymphocytes or of
dialyzed TF are set out. The follo~ving figures are from that table and indicate
the proportion of subjects, all initially nonreactive, who gave positive skin
reactions after treatment:
Sarcoid 11/13 Cancer 21/43 Kwashiorkor 12/12.

In contrast, patients with Hodgkin's disease given leukocyte injections showed

0/42 conversions, as compared with 41/43 control subjects. There are occasional
suggestions of clinical improvement in the conditions showing appearance of
skin reactivity, but most are unaffected. The appearance of skin sensitization
in malignant disease is often of low level, much better seen near the site of
lymphocyte or T F injection and disappearing more rapidly than in normal
recipients.
The use of T F in persisting infections associated with apparent paralysis
of T immune response has been extensively tested within the last two years, and
there are earlier reports of the use of donor lymphocytes that can perhaps be
assumed to work on the basis of TF. As yet the results seem to have no special
bearing on the interpretation of T F activity and no attempt will be made to
assess these reports.

THEORETICAL ASPECTS
We can begin a brief, and necessarily rather superficial, discussion of the
biological nature of Lawrence's transfer factor by using Lawrence's own sum-
mary of the nature of the phenomenon. ~ The molecular nature of the active
agent is unknown. The filtered enzyme-treated extracts that are referred to as
T F contain a number of components, and it is suggested by Lo Buglio is that
only one of these is active. Lawrence regards it as established that both
nucleotide and peptide units are present in T F and hints at the possibility of
there being a short, double-stranded RNA. T F could, in his view, still represent
an informational molecule conveying immunological pattern to the recipient
cell. However, Lawrence appears now to favor the alternative that T F acts as
a de-repressor of normal lymphocytes, or, more correctly, of "a select small
population of normal circulating lymphocytes," which, after exposure to TF,
become antigen-responsive, transform, and undergo clonal proliferation.
It is self-evident that the most urgent research need at the moment is to
6 Burnet J. ALLERGY CLIN. IMMUNOL.
JULY 1974

identify the chemical nature of the active agent and the way in which specificity
in relation to the various standard antigens is expressed at the chemical level.
I n view of the small molecular size of the agent, it is impossible to equate it
with T receptor substance that, following Marchalonis, Cone, and Atwell, 17
can be accepted as an immunoglobulin. Equally, there is on the available
evidence no possibility of its being an antigen or a macrophage-processed "super-
antigen." Some new category of information-carrying molecule must be
postulated.
Lawrence has already foreshadowed the general f o r m that discussion of the
T F phenomenon will t a k e - - t h a t it represents either the t r a n s f e r of an informa-
tional molecule from donor celt to recipient or that it is a trigger or de-repressor
allowing the recipient cell to synthesize and liberate molecules or complexes
of like character. There are clear indications that the T F process subserves an
important biological function, and I am strongly influenced by the dogma that
adaptive genetic change, whether in the germ line involving the whole organism
or in somatic cells, is never "instructively" induced by environmental impact
but results always from the action of selection on phenotype modifications arising
from random genetic processes. I f c u r r e n t theories of the genetic control of
protein synthesis are correct, TF can act only by stimulating recipient lympho-
cytes to produce more TF. It is as much a product of the stimulated cell's
genome as antibody or a histocompatibility antigen.
The simplest way to visualize the process is to accept that T F (whether
molecule or complex) can recognize a minority of lymphocytes whose phenotype
makes them capable of synthesizing T F of similar pattern. Obviously, if T F is
to recognize and stimulate a T cell of the right type, there must be a cell s u r f a c e
receptor with which it can react, inducing an appropriate signal calling for
cellular activity. H a v i n g regard to the small molecular size of T F , the com-
plementary receptor could be spoken of as a minireceptor (MR). Superficially
at least, the T F - M R specificity seems to be roughly equivalent to the broadest
reactivity of the fully accessible T cell-immune receptor ( T R ) . This a p p a r e n t
correspondence of T F - M R specificity with that of the IgM-based immune
specificity of the T cell is probably the main theoretical difficulty in providing
an interpretation of TF. The T F - M R relationship is formally equivalent to that
of antigenic d e t e r m i n a n t / a n t i b o d y combining site. No one seems to have dis-
cussed the existence or nature of the MR, but it seems as unlikely that they are
immunoglobulins as that the T F ' s have any molecular resemblance to the
antigens to which they seem to be related.
A schematized attempt to hypothecate the nature and origin of the postulated
T F - M R system could be as follows :
1. T F and MR are small macromoleeules with mutual steric relationships.
broadly equivalent to AD and to CS of antibody or immune receptor, respective-
ly. Both are coded for by the cell genome and both are, presumably, associated
with the cell surface. MR can be visualized as a specific configuration of some
maeromolecule forming part of the cell membrane.
2. The development of a limited diversity of T F patterns to aceount for the
observed degree of specificity is assumed to be analogous to the still unresolved
VOLUME 54 Transfer factor---a theoretical discussion 7
NUMBER 1

FIG. 1. Relationship of TF specificity and immune specificity in a differentiating T cell clone.

I, Level at which TF specificity is expressed and stabilized. Each TF pattern is shown by a
number: 22, ,$7, etc. II, Level at which cells of superclone 57 express a single specific
immune pattern: AAB, .Jllu~A, etc. It is suggested that most or all of these immune patterns
will have some cross reactivity.

genetic process by which antibody diversity is generated. It could well be a

persisting representative of the earliest stage of the evolution of specific im-
munity. It would be an essential part of the process that, once a stem cell had
reached the appropriate stage of differentiation, it and its descendants should
express only one type of TF.
3. It must be postulated that the steric patterns by which MR and TF react
and "recognize" each other are of complementary structure, the mutual relation-
ship being coded for by the cell genome.
4. Production and functioning of TF-MR is confined to immunocytes and
their precursors, but the assumption is made that the TF-MR pattern is fixed
before, perhaps many cell generations before, the specific antibody pattern is
stabilized and phenotypically expressed.
As a model system schematically represented in :Fig. 1, we may suppose t h a t 100 TF-MR
patterns are produced and t h a t at least one of each is represented in the first 200 stem cells
destined for T immunocytes. I f we take a single cell with p a t t e r n No. 57, as this proliferates,
the c0ntinuing processes by which generation of antibody diversity (VL-VH) is effeeted will
 Burnet J. ALLERGYCLIN. IMMUNOL;
JULY 1974

~Ag

TR MR TF
producer

Ag LK TF LK

]I
TR
MITOSIS

MR

Ag TF 0 O LK

MR
MITOSIS
FIG, 2. Schematic figure of TF hypothesis. I, Unstimulated T ceil surtoEe wire tmfY~une
specific TR and NIP.. A potential TF pNxlur (rrlocromolecuJe or orgonet|e} Js indicated.
II, Stimulation by antigen induces increased membrane permeabi.lity with lymphok~e
(LK) release, active TF production, and miteiis. I!1, Stimu|ation by TF induces increased
membrane permealmlity with lymphokine release and active 11= production, but no mitosis.
IV, Stimulation by TF and A9; all systems activated.

go on, allowing another 100 antibody patterns to emerge m "superclone 57" bofore pheagtyt)ic
restriction becomes effective. On any theory of diversification, the immune pattemas of "57"
descendants will have some degree of mutual resemblance.
Most clones, for example, which have immune T type receptors (TR) reactive with PPD
will be from a single superclone with a uniform TF-MR pattern.
As long as the processes follow the rules given, there is no reason at all why ~he M,R's
should not have a totally distinct molecular structure from that o~ the TR on the same celt.

5. I t seems i n e s c a p a b l e t h a t a l l p a t t e r n - p a i r s o f T F a n d M R Should be c o d e d
b y g e r m - l i n e genes. I t m a y be d e s i r a b l e to e x a m i n e t h e p o s s i b i l i t y t h a t t h e m o u s e
locus I R 1 , a n d c o r r e s p o n d i n g genes in o t h e r species, m a y b e a s s o c i a t e d w i t h t h e
T F - M R system.
6. A t t h e e v o l u t i o n a r y level i t h a s b e e n s u g g e s t e d t h a t one c o u l d d e r i v e
mutually reactive patterns from the need for recognition of the appr~ate
q u a l i t y o f m e t a z o a n cell s u r f a c e s in contact. ~s I f A / B r e p r e s e n t s t h e p r i m i t i v e ,
m u t u a l l y r e a c t i v e p a i r , one m i g h t s u g g e s t t h a t in d u e c o u r s e A c o u l d g i v e rise
VOLUME S4 Transfer factor--a theoretical discussion 9
NUMBER I

to TF's and histocompatibility antigens, while B was the prototype of MR, the
immune receptor on T and B cells and the immunoglobulins.
7. It will be noted that, despite the correlation with immune reactions, the
TF-MR system is not directly reactive in any way with the antigens concerned.
This concept of the nature of the TF-MR relationship can now be applied
to a description of what is happening in the various reactions involving TF
(Fig. 2).
Unstimulated (naive) T cells, recently differentiated in the thymus, are
assumed to have some MR's presenting on the cell surface, but probably no TF's,
or at least none that can be liberated. Specific TR's are few and relatively inac-
cessible to any but "best-fitting" antigenic determinants. When a potentially
sensitizing antigen enters the body, say as a focus of tuberculous infection,
antigenic fragments of various types will soon be in circulation. Interaction of
antigen with the immune receptor (TR) of T cells of appropriate reactivity,
i.e., ARC's, will eventually occur. Such reaction is postulated to have two effects
of major relevance in the present context: an increase in the number and ac-
cessibility of the antibody-like TR's and the synthesis and appearance on the cell
membrane of TF's and perhaps of increased numbers of MR's.
Once this has happened in an adequate number of cells, some of the newly
synthesized TF's will be liberated from the cell surface and will find other naive
ARC's suitable for attachment. The basic hypothesis of TF action is that union
with a naive ARC of appropriate specificity will act as a signal for TF synthesis.
There follows an autocatalytic process that will result in a large proportion of
ARC's potentially reactive with tuberculin becoming mature sensitized cells. It
is assumed that the effect is essentially what Lawrence 7 found with his in vitro
experiments, using normal lymphocytes, TF and PPD. If, for the time being,
we forget about proliferation of sensitized cells, it is obvious that a process of
this sort would facilitate the rapid mobilization of all existent cells that had a
specificity that could include reaction with tuberculin. It is a concept that seems
to fit well with other bodily methods of handling an emergency, such as the
cascading activation of the various factors concerned in blood coagulation.
The real test of such an hypothesis takes three forms. One is its heuristic
value in suggesting experimental methods by which various points could be
tested or a molecular interpretation of aspects of the process provided. The
second is its capacity to make sense of what happens in immunodefieiency
diseases or other pathological conditions and suggest new methods for investiga-
tion or treatment. The third is to see whether it "makes biological sense"--
whether such a mechanism could be evolved, and, having evolved, would provide
increased survival to those forms that had developed it. Included here will be
the need to interpret the apparent conspicuousness of the mechanism in man
as compared to other mammals.

CONSEQUENCES OF THE THEORETICAL APPROACH

Experimental implications
As has been mentioned more than once, the first requirement for more
effective understanding is the molecular identification of TF. Since
ribonucleotides are known to be present in all active extracts, most authors have
10 Burnet J. ALLERGY CLIN, /MMUNOL.
JLJLY 1974

favored an RNA of some kind. Having regard to the low MW of all components
in DTF, the only types of RNA that could be considered are soluble transfer
RNA ~70 to 90 nucleotides, MW 21,000 to 28,000), some disintegration product
of it, or some other RNA. Clearly, chemical study of T F will eventually call for
the cooperation of biochemists skilled in the chemistry and enzymology of
transfer RNA's.
In this connection it is of much interest that recent work in plant pathology
(summarized in Science 178: 734, 1972) has found very small infectious RNA
molecules, with a general resemblance to s-RNA, responsible fer the transmissible
plant diseases, potato spindle tuber, chrysanthemum stunt, and Citrus exocortis.
These are now being spoken of as viroids. This raises the thought that any
autocatalytic system of the type being postulated for TF-MR in human
lymphocytes, with transfer of an active molecule from one cell to another, must
surely have occasional possibilities of pathological mutation to a viruslike
activity. There have already been suggestions that some of the more puzzling
human virus diseases, such as kuru and hepatitis B, may have some related
etiology.
Despite all this, it has still not been proved that TF is RNA, and the nature
of MR has not even been considered by anyone.
It is inconceivable that the TF phenomenon, as seen in man, is not present
in other mammals in some form, and it is important to look both for its
manifestations and for any substances or processes that render it more difficult
to demonstrate in experimental animals than in man. There are already
persuasive claims to have demonstrated the phenomenon in rhesus monkeys and
rats. The biological interpretation of TF, as part of a cascade mechanism to
mobilize defense processes rapidly, calls for further study of the lympl~okines,
which have been assigned a similar function. It would probably pay to under-
take a repetition of lymphokine experiments, using dialyzed materials and
checking for specificity according to antigen, at appropriate points in the study.
A reported finding that is not easy to fit into the pattern of the hypothesis
that has been adopted is the sharp specificity in the transfer by TF of capacity
for second set rejection of homografts.19 If both hypothesis and observation are
correct, their mutual adjustment might be highly relevant to Jerne's concept
that the basic patterns of antibody transmitted in the germ line are solely
concerned with histocompatibility antigens characteristic of the species. In view
of the difficulties of reading rejection times and the possibility of subjective
bias, any repetition of such experiments should be done triple blind, with neither
the surgeon, the experimental recipient, nor the person reading the results aware
of the grafts' origins until the experiment is completed.

Immunopathology
There are a number of interesting qualifications of such a theoretical ap-
proach to be drawn from, perhaps, the best studied of these pathological situa-
tions: chronic candidiasis.2~ In a typical case there is ample production o~
antibody and the patient's lymphocytes sometimes respond to P t t A or Candida
antigen by DNA synthesis but never by lymphokine production. There is usually
a development of capacity to produce MIF after TF, but no change in the
VOLUME 54 Transfer factor--a theoretical discussion 11
NUMBER 1

usually negative DNA activation results. The essential effect of T F is therefore

not in inducing cell proliferation but in conferring or restimulating the capacity
to liberate lymphokines, the essential agents in cascade effects. It still remains
to be determined whether T immunocytes are monofunctional, i.e., composed of
separate populations (1) producing lymphokines with the further reservation
that there may be subpopulations, each with a limited repertoire of one or more
lymphokines, and (2) cells capable of proliferation on specific stimulation
(memory cells). The alternative that any T cell in normal condition can show
both (1) and (2) responses on stimulation is not yet excluded.
It seems most unlikely that chronic candidiasis is ever seen in an im-
munologically normal individual, and we can assume provisionally that there is a
genetic defect in the sense that specific stimulation of T receptors does not provoke
T F synthesis, and perhaps concomitantly there is failure both of lymphokine
production and, less regularly, of ability to induce cell proliferation. This could
also be associated with the tolerigenic action of excess antigen in eliminating or
inhibiting all adequately reactive clones of T cells. Administration of T F would
have, as a primary effect, stimulation of any still extant cells with appropriate
MR's to synthetic ability, producing, first, TF-MR pairs and T receptors, and,
concomitantly, MIF and other lymphokines.
All the conditions that T F can to some extent ameliorate or correct seem
to represent failure of responsiveness of lymphocytes to varying degrees. Many
are clearly genetic, but one can also conceive of metabolic conditions that could
reduce the reactivity of genetically normal immunocytes. The well-established
nonreactivity of grossly undernourished children is an obvious example, is and
another likely case is that of Hodgkin's disease. 14 Here the anergy appears
early in the disease and on occasion may revert to normality with a remission
of the disease. Depending on the degree or type of the functional deficiency, the
addition of T F may, at one extreme, provide an autocatalytic trigger toward
normality, while in others, of which Hodgkin's disease is the classic example, it
is unable to produce any significant effect. Any perusal of the published protocols
will show many differences and anomalies. In particular, the more refractory
intermediates tend to show a transfer of skin sensitization only at the local
site, and, if a systemic effect is obtained, it is a much more transitory one than
in the standard experiment with healthy young adults.

Biological significance of transfer factor

The gist of the hypothesis that has been outlined is that transfer factor is one
of a series of mechanisms by which a bodily emergency can be rapidly dealt
with. Specifically, it is part of the process concerned with the mobilization of
T immunocytes. In the normal individual with no significant previous experience
of, say, tuberculosis, there will be cells of several clones that can react with P P D
or other TB antigens, and a small proportion may have come from cell lines that
had experienced and reacted to some related antigenic determinant. Most, how-
ever, will be present as naive immunocytes (antigen-reactive cells) carrying no
T F and with specific receptors relatively inaccessible and likely to be stimulated
only by an accurate high affinity contact by antigen.
With the appearance of an infection or some other emergency, an initiating
 12 Burnet J ALLERGY CLIN. tMMUNOL.
JULY 1974

cancer, perhaps, foreign antigens enter the lymph circulation. Soon small num-
bers of T cells with appropriate receptor patterns are stimulated b y antigen
contact. Once this happens, a cascade effect develops, resembling in some ways
that described by Macfarlane 23 for the blood-clotting sequence. As in that
situation, one can feel sure that the process of lymphocyte mobilization also
possesses its in-built feedbacks and fail-safe mechanisms. All will provide chances
for genetic anomalies to occur and manifest their phenotypic expression in some
degree of immunological inadequacy.
The cascade process presumably begins with the stimulus to synthetic activity
provided by antigenic contact with a T cell immune receptor. The resulting
activation means that more monomeric IgM receptors are synthesized and are
more accessible in relation to the cell surface, and concomitantly T F units
appear on the surface. Subsequent effective contact of the cell ~ i t h antigen
induces, by hypothesis, two distinct processes. A proportion of T F ' s are liberated
as free molecules or complexes into the lymphatic circulation and the IgM
receptor develops a wider range of specific reactivity. B y hypothesis, the
liberated T F units will find it possible to attach to and activate any T cells
whose MR's (perhaps we can speak of them as " T F sockets") are of appropriate
pattern. Such cells, in their turn, synthesize and liberate T F and develop more
accessible and more widely reactive Ig receptors. In the course of this widening
process of stimulation, lymphocyte proliferation and liberation of lymphokines
of various sorts will occur. It is not known how the capacity for and the occur-
rence of these reactions are distributed through the lymphocyte population.
Also. it is by no means clear how m a n y lymphokines there are. nor the n a t u r e of
those that appear to be able to stimulate previously quiescent lymphocytes to
activity. One cannot exclude the possibility that T F is at least in part responsible
for some of the activity that has been ascribed to lymphokines
The over-all result is that the small initial population of specifically reactive
T lymphocytes is rapidly reinforced by a large population of activated lympho-
cytes, all of which are available for concentration on an infe~tious focus or
to give the erythema and induration of a positive skin test.
Clearly, the phenomenon makes good biological sense if looked at from this
point of view. and I have no doubt that in due course it will be possible to
provide a credible story of how this particular lymphocytic function evolved.
One feels, however, that until the chemical nature of T F is known, any evolution-
ary discussion would have to use such generalized concepts that the result would
have no intellectual elegance whatever. And without some degree of elegance,
evolutionary discussion is of very little interest to anyone.

ADDENDUM
I n a c u r r e n t article ~ M a r c h a l o n i s , Morris, & H a r r i s e l a b o r a t e an e s s e n t i a l l y equivalent
a p p r o a c h as t h e m o s t f a v o r e d o f several a l t e r n a t i v e h y p o t h e s e s to accoun~ f o r t h e influence
o f I R g e n e s on T cell r e a c t i o n s in mice, a n d p e r h a p s f o r the a s s o c i a t i o n of t h e m a j o r h u m a n
h i s t o c o m p a t i b i l i t y a n t i g e n , H L - A S , with several i m m u n o p a t h i c conditions.

~Marchalonis, J. J'., Morris, P. J., a n d H a r r i s , A. W . : I n press~ J. I m m u n o g e n e t i c s . 1974.

VOLUME 54 Transfer factor--a theoretical discussion 13
NUMBER I

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