CA2129895A1 - Autoantibody assay and usage in the control of human disease - Google Patents

Abstract

Methods of assay for and therapeutic use of autoantibodies, specifically anti-thymocyte autoantibody (ATAA). This is a teaching of methods for the assay of autoantibodies and for ATAA use, based upon autoantibody quantity in relation to a target lymphocyte population, for predicting body resistivity/susceptibility to disease and tumors, as well as suggesting protocols for the treatment of various diseases, malignancies and autoimmune reactions. The main thrust of the disclosure is the inventor's thesis that, by altering the amount of autoantibodies (in sera) which regulates the target cell population, diseases and/or malignancies may be eliminated or otherwise controlled.

Description

212'~
W~ 93~157~g PCr/lJS93/1)0886 AUTOA~TIBODY A5SAY AND USAGE IN THE CONTROL OF
HUMAN DISEASE

BP~CKGROUND OF THE I~VENTION
1. Field of the Invention This invention relates generally to the field of biological cellular antigen assay and, specifically, to establishing a method of assay for autoantibodies in relation to regulation of the immune system. This includes, and herein teaches, a novel method for relating : ~ the assay to predict body reslstivity (or susceptibility~
to autoimmune diseases and tumors, thereafter suggesting a pro.tocol for treatment of the aforesaid diseases by increase/reduction of the autoantibody titer.

~ ~ , 2.~ Discussion of Relevant Publlcations -;
All ~species tested ~rat, mouse, man etc.3 have a : demonstrable ant1body (ATAA) in their serum which is capable ~oP ~eactlng with and sometimes killing ( in vi tro) a:~selected population:of their own thymocytes ~autologous~
or thymocytes of~ the same species (allogeneic). This antlbody ~ill also react in certain species with bromelain-treated red blood cells and possibly dendritic epithelial ~;
cells, CD5 receptor and a presently unknown receptor specific fQr ATAA. During the last decade or two, various ;investigators have deta~led a wide ~ariety ofautoantibodies l~pecific antibodi~s which react with almost any tissue ~r or~an in the body, i.e. brain, skin, various serum proteins ( lncluding antibody molec~les ), all blood s WO93/15759 : PCT/US93/00886 cells (including lymphocytes), intestinal tissues, heart cells, etc.]. It has never been clear whether these antibodies cause resultant disease or are the result of the disease, but in some cases they are interpreted as being causal. ~:~
One swch autoantibody is one directed against antigens on thymocytes (ATAA). There are many different types of :
thymocytes which originate in the thymus and are the precursors of T-lymphocytes which are found in blood and various lymphoid and non-lymphoid organs of the body. T-lymphocytes are concerned with cellular immunity, as differentiated from B-lymphocytes which produce antibodies and are responslble for the humoral immune response. The T-lymphocytes recognize antigens by recognition sites and accessory molecules which are of various types and which define each type of T-lymphocyte. The recognition sites ,, are associated wlth :the ma,or histocompatibility complex.

; Thus/: T-lymphocytes are of many different types, each :
:iden:ti~iable by distinct antigenic differences and many of which have distinct functions. The development ~ontogeny) of the various T-lymphocytes occur within the thymus gland and are present ln various percentages, at various times, within the thymus. ~(See: Molecular and Cellular Events of T-Cell Development; B.:J. Fowlkes and Drew M. Pardoll; Adv.

.
In Immunology; Vol. 44, pp. 201-217; 19891.
Aside from classification of T-lymphocytes by their recognition sites and accessory molecules, there aré
several different functlonal subsets of T lymphocytes:

2 1 2 ~ .) J
15759 P~T/US93/U0886 :
helper cells; suppressor cells; killer cells; etc.
therefore, it lS no surprise that investigators have been unable to understand the nature or the functional significance of the thymocyte autoantibody. It has been said thàt the serum levels of ATAA do not show etiological significance to disease (Eisenberg et ~l., J. Immunology 122, p . 2272, 1979 3 .
MeasuremPnts of ATAA are made in vitro in which antibodies in serum are mixed with an animal's thymocytes and.~a reaction~is~observed~which can~lead to the binding of and/or death of a certain number of the cells. It is .:
llogical to think that~such antibodies and target cells could coexist in~vi~w. Thus, it is not surpr1sing that a disease causal effect has not been postulated. There has :recently been des~cribedi in certain strains of mice (motheaten ~ and NZB),~; ~a ~rapidly fatal condition :characterized~by a:wide variety of autoimmune diseases and , a:n~ extraordinarily::high level of ATAA. It is of special interest that ~some of :these animals have naturally occurring anti-tumor activity.
~ In~the:same vein as the above, Perper et al. disclose ;~ an IgG~ thymolytic autoantibody in rats which has : specificity for a sub-population of T-cells (R.J. Perper, :: A.L. Oronsky and Maria~Sand~ mmunology, paper 312; 1976)-.
: ~ These researchers present an interesting disclosure wherein a cyto~oxic an~i-thymocyte IgG autoantibody is found .
;~ present in Lewis rats which, in the presence of autolo~ous complement, destroys (in vitro) 12-Z8~ o~ isologous or - :
:

~2~'~9~
WO ~3/15759 . PCT/U~g3/00 ~utologous thymocytes, a small number of lymph node cells and splenocytes, but not bone marrow or circulating lymphocytes. The labile cells in the thymus represent a finite subpopulation which is autologous anti-thymocyte antibody sensitive and steroid resistant. The presence of the autoantibody is randomly distributed in outbred animals, whereas inbred Lewis rats, a strain in which the induction of some autoimmune reactions is under genetic control, the antibody is almost always present. In this strain, the susceptible T-cells and the quantity of circulating autoantibody is significantly depressed during the productive phase of the T-cell mediated disease (adjuvant polyarthritis~ and returns to nor~al after the disease becomes stabilized. There is a direct relationship seen between~the amount of susceptible cells in the thymus and the amount of~antibody in circulation, which suggests that the antibody could serve as a marker for a specific subpopulation of thymocytes which may have a regulatory nfluence on T~cell reactivity. But Perper et al. stop at this point and, the aforesaid disclosure notwithstanding, recent patent and~ professional literature searches have failed to reYeal a teaching of what I now term ATAA
causality. !
The concomitant presence of both ATAA and autoimmune .
diseases in the same animal continues to frustrate other investigators because none have yet been able to establish a causal role for ATAA, simply because they are making measurements of ATAA in the presence of disease. .

2 1 2 ~ 9 ~) W~93/1~759 PCT/US93/00~X6 Interestingly enough, U.S. Patent No. 4,937,071, teaching a METHOD FOR AUGMENTING IMMUNE RESPONSE, discloses a method for enhancing the ability for humoral immune response in a -;
mammal which comprlses exposure of lym~hocytes `~
': .';
: histocompatible with the lymphocytes of the mammal to the ~ ~ .
presence of del:ta-~immunoglobulin at a concentration higher than that at which the~ lymphocytes~would have been exposed while in the l:ymph or blood ~stream of the mammal an~, thereafter introducing these lymphocytes to the blood stream:of lymph of~the mammal. This is clearly a method of : augmentation,~and:no one~suggests the use of a naturally occurrlng (or~monoclonal~replicate of an) autoantibody to re~ulate~the i = ~response mechanism. This fact exis.ts in~spite of the~:~proliferation of a great deal of related ~ork;~such~as :the~preparàtion of monoclonal antibody for :inhl~itlng adhes`lon~-d ndent l~eukocyte f;unctions, the use of~ monoclonal~antlbody~ for;~:~diagnostic procedures use~ to ;di~ferentlate~ between~ normal~ cells and~ tumor cells diagnostic:~:and~th~erapeut~ic~uses~of~a~monoclonal antibody ag t the~anti ~found on~essentially all~normal T-cells and:~ cutaneous~:T-l:ymphoma cells,~ the identification of monoclonaI anti~-S-~antibodle~ and:the di~sclosure of varlous com`pounds~ ~hat~ :are~ sald~ to modulate immune ~response ib3d . ~ rurther~to~this abundance~of~antlbody~llterature, Harley;l in ~U~5;.~ Patent No. ~9,784:,942, ~teache~s the .production: of ~ono~lonal:~ antibodies,~ produced by a continuous :hybridoma~cell l~ne~ in~methods~ for dete~tin~

the presence: o~ s;elected~autoimmune RNA proteins and 21~9~
WO93/15759 P~T/US~3~00$~

. .
antibodies against such proteins in biological samples~ and which may be incorporated into diagnostic test kits for the slmilar purpose. The monoclonal antibodies used against autoimmune RNA proteins include proteins such as La~ssb, Ro/ssa, aNP and Sm. These monoclonal antibodies may be applied in methods for screening subjects for systemic ~, lupus erythematosus, subacute cutaneous erythematosus, neonatal lupus, Sjogren's syndrome, complete congenital :
heart ~lock, and~other disorders which involve the presence .~
~ of antibodies against autoimmune RNA proteins. In spite of ~
~ . :
painstaking searches in the literature of the U.S. Patent Office and the profes;sional journals, I have found no , reerence,~ of any kind,~to the~inculcation of a predictive value for imputing the~ ~usceptibility for autoimmune di;seases.~
The~ above~mentloned shortcomings which appertain ~;
within the study~o~f~autoantlbodies and their relationship t~o ~ carc~inomas and ~autolmmune~ diseases have been s;atisfact~orily~answered by~my research and are reveal~d in `;
he~herelnafter~dlsclosed assay and treatment methods.

SUMMARY
have found~quite surprlsingly that the target cell ;of ATAA regulates~the~ development of the disease and the ~;
level of ATAA serves as a marker and may control the number `;

of these (target)~cells. I use the amount of ATAA to predict susceptibility/resistivity to disease ~nd to regulate the guantity of these target cells so as to be : .

~ '".. ' 2 1 2 ~t;~.Çs.~J~
WO93/1575~ PCT/US93/00886 able to manipulate body susceptibility/resistivity and the cause of both autoimmune diseases and malignancies. The quantitation of ATAA in circulation using the appropriate target cell (a subpopulation of thymocytes, the antigen of which might be present on various other cell types and detailed, but not limited to those mentioned earlier) predicts susceptibility to the development of allergic hypersensitivity reactions and autoimmune disease or resistance to tumor development. Furthermore, by either raising or lowering the level of ATAA, or the target (susceptible) cell, modification may be had of the course of either tumor development or autoimmune disease/a~lergic hypers~nsitivity dise~ses.

: :: : :
BRIEF ~DESCRIPTION OF THE DRAWINIGS
Of the drawlngs:~ ~
Flgure i~ l~S a graphical representation showing the :effect ~of ~pre-existing autoantibody titer on the severlty:of adjuvant dlsease~(re: swelling) 1n Lewis rats;
Figure~ 2 : lS a~ graphical representation ;:contrasting~ the level~ of pre-existing ~anti-thymocyte autoantibody (ATAA) with the change in b~dy weight ~day 8-: ~ 18) after the induction of EAE; and ~:: :- Figur 3 is~a graphical display which illustrates .
:~ :thatt when the target ~cell ~of ATAA is eliminated, there exists an enhanced immune reactivity of the surviving .:
~ cells. ~

`,'~

~12g(,~ ' WO93~15759 PCr/US93~008B~

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT
The target cell of ATAA is able to suppress the cellular immune response (a suppressor cell in, and derived from, the thymus). High levels of ATAA reduce the number of suppressor cells and allow the development of autoi~mune diseases. Alternatively, low levels of ATAA result in high levels of suppressor cells which prevent cellular immunity from rejecting malignancies~ Thus, an inverse relationship .

between~ vulnerability to mallgnancy and autoimmunity exlsts. Thls relationship is regulated by the presence of ATAA. The reason that serum levels of ATAA do not correlate with the progression of disease is because, when an~autoimmune disease appears, there may occur a production and release from~the thymus~ of suppressor cells in an attempt to abort disease development. These cells consume circulating ATAA which ~results~in a drop in its level in cir~ulation. Thus~l;;the (conventional) measurement of ATAA
durlng ~disease~ ~will~produce anomalous data that are completely misleading. ~ ~
The~inductlon of~adjuvant arthritis in rats has been shown in multiple~published studies to be a model of animal and~human autoimmune; disease, as has the induction of allergic encephalomyelitis (EAE). In the arthritis model, , ~: : ~ , the amount;~of~AIAA (as measured by the ability of serum to ~;
kill a~ subpopulation~of thymocyt~s --see description of assay hereinafter) predicts the severity of the arthritic condition (Figure 1~). When EAE was induced in rats, the :: .. .
pre-existing quantity of ATAA predicted the development of ':

W093/1~7s9 ~ PCT/US93/~0886 disease as quantitated by the change in weight during the disease process (Figure 2). Therefore, when an individual has a high amount of antibody, he/she is more susceptible to the development of autoimmune disease induction. A
reasona~le assumption is that the antibody destroys a population of thymocytes which repress the development of the disease; and when a ~stimulus is applied, disease deYelopment runs unchecked. This is best exemplified by using another model system in which a suspension of foreign cells is injected into a rat. These cells are so constituted that they can react against the recipient, bu~
the recipient is~unable to react against the cells (graft ~ersus ~host-recipient~ reactionl. Th~ reaction is quantitated by measuring the size o the lymph nodes draining the injectlon site. When :cells are treated with ATAA,~there ls a greater than~expected reaction ~Figure 3) which-lndicates ATAA~destroyed a cell which is capable of suppresslng the reactlon, thus resultin~ in a greater than expected (predic~able3 activity. Thus, Figure 3 shows the ::: : : :
: popliteal lymph~nod~e Graft ~ersus Host (GVH) of Lewis rat ~;;

thymocyte:s preincubated with isologous serum (naturally , occurring :~TAA~ The ~same volumes~ of cells at varying concentrations were~injected~in the ~ub-plantar location of F.Hybrid~(L/BN).~ Thymocytes were preincubated with either comp~ement~sufficient isologous serum (120 CH50 units) or the same serum heat inactivated (0 CH50 units~. The control~:
, experiment in ~which 5~0 x 107 F.Hybrid cells were ~:

: preincubated with complement sufficient is shown as a ,.

~.

212~5 WO93/15759 PCT/US93/00~86 single point (L/BN L/BN).
The reason that ~ther investigators have missed the prognostic significance of ATAA and its relationship to disease is because they measure the levels of antibody while ~he disease is present. However, it is revealed through my studies, that after the induction of an autoimmune disease in rats (adjuvant arthritis), the susceptible cells as well as the amount of ATAA decrease markedly. Thus, any measurement of these parameters while (an actlve) disease exists would preclude any conclusion concerning etiol~gical significance. This observation further helps to explain the presence of ATAA, in c~irculation, when the target celI is within the same body.
Sin~e there h~s been reported to exist a functional barrier between cells within thymus and antibodies in circulation, :i:t:~is reasonable that:ATAA exists in circulation coincident w~th target cells with tbc thymus. When, however, cells are:~released (due to lnduction of disease or due to genetic programmi~ng) the reaction between ATAA and target cells resu;lts in a permanent decrease in numbers of susceptible cells and a ~temp~orary~decrease ln ATAA. Once the susceptible cells are eliminated,: the amount of antibody increases, since lt is still being produced. Now, if cells were~slowly released, they would be destroyed; but, ATAA
would remain high, as~ we~have previously shown that ATAA
exists in excess in~circulation.
Although ~I lnculcate a theoretical role for the regulation of the immune:response by ATAA, ~here is a~so :

WO93/15759 21 2 ~ g ) ~, PCT~US93/00886 the empirical datum that the production and presence of ATAA is controlled or linked to the same immunoregulatory gene (Ir) as that which causes the disease itself; i.e., one rat ~ets disease whereas another does not because of its genetic makeup and the expression of this gene is accompanied by the expression of the gene which results in AT~A production. In either case, however, I have discovered that the measurement of ATAA production and/or the target cell antigen(s) of ATAA has diagnostic, :
prognostic and/or therapeutic value. ~:

I apply the same type of assay system as has been used .:
in rats to a human situation. In this case, I use various human sera and fetal human thymo~ytes as target cells.
Using this assay, I measured, in several human samples, the presence o ATAA which reacts with (kills) a small percentage of human fetal thymocytes. There was variability in the number of target cells killed between various human samp1es, including one in which an aberrant immune responsiveness ~existed historically. As mentioned earlier in the SUMMARY, the quantitation of ATAA in circulat1on while~ using the appropriate target cell, predic~s~ susceptibility to the development of allergic hyperser1sitivity reactions and autoimmune di~ease ~high level) or resistance to tumor develQpment (low level). By either raising:or lowering the level of ATAA or the susceptible cell, modifica~ion of a course ~ either tumor de~elopment or autoimmune disease tincluding aller~ic hypersensiti~ity diseases) is attained.

, 2 ~2g'~39'~ , W093/15759 PCT/US93/OOB~-.

There are other uses for an assay using ATAA and relevant target cell or target antigen. This is detailed in the following table:
Table A
UTILITY OF ASSAY (Regulatory Relationship of Antibody to Target Cell Antigen) The assay permits:
1. The prediction of vulnerability ~or development of a~toimmune diseases so that patients can be tested at regular int~rvals for the appearance of autoantibodies of various types. Treatment may be s~arted early. These data may be used for genetic counselin~. ;
: 2. The prediction for drug idiosyncratic reactions since many are autoimmune i~ origin.

3~ The prediction for susceptibility to reactions to immunizations or any other allergic hyper~
sensltivity reaction.

4. :Use as a marker for following the efficacy of ; ~
. ., ~arious trea~ments of an autoimmune disease.

5.~ A reference as necessary to monitor efforts to :: ~: . .
alter levels of ATAA and susceptible cells as .-: ~ outlined further in Table B.

6. The prediction ~ulnerability for development of tumors so that patients receive enhanced medical surveillance for early tumor detection, and thus, alerts a patient to avoid high risk activities. -It is ~lso important for genetic counseling W~93/1575~PCT/U~g3/0~886 concerning tumor development.

7. The monitoring of course and treatment of patients with tumors.

8. The use for development and assay of new therapeutic agents for treatment of tumor and autoimmune diseases. (drug screening tool).
.:

9. The use to design new therapeutic agents which :

c~n modify ATAA or its susceptible cell.

10. Deletion of self reactive cells a~d/or suppressor . cells.
..

11. Retardation of transplantation rejection.

12. The enrichme~t of suppressor cell population in .~,..
bone marrow graphs in order to prevent graph versus host reactions.
: Table B outlines methods for altering the in vivo levels of ATA~ or the susceptible cell. This is simply an .
outline without excludin~ other possibilities currently -;
nown or which~:may~become available, and the reader should be cautioned not ~to attribute arbitrary bounds to these .
: methods~

: Table B
. ~, METHODS OF A~TERING LEVELS OF ATAA AND SUSCEPTI~LE

:: : OE LLS AND VTILITY~FOR SAME
:
I ~91~lr9LL9Y~I~LY~_~3~

A. Passively transferring serum from donor with : ~
high levels of ATAA~

B. Production and transfer of monoclonal ATAA~
`

~1 ~9~3 WO93/1~759 PCT/US93/008 C. Use of immuno stimulants.
UtilitY
1. Enhance tumor rejection.
2. Prevention of tumor development.

': .-, II. Lowerinq Levels of AT~A
A. Passing patient' 5 serum (extra-corporeal) oYer antigen co~taining immuno-absorbents.
B. In vivo administration of purified target cell antigen.
C. Administration of pharmacological agents ~which are capable of lowering existing antibody levels (as yet to be described).
U~i~it~
1. Prevention and/or therapy of autoimmune dise~se. ~--..;-,. ~:
2. Prevention of hypersensitivity reasti~ns.

. Increasinq_Number o~ Susceptible Cells ~-~ .
A ~ Lower levels of ATAA ~ see abc)ve ) .
B. Adoptive transfer from fetal or adult ~;
source.
C . In vi tro: delete other lymphoid ~:ell types resultlng in functional increase in susceptible cell.
tility 1. Same as lowering ATAA -- in vi vo ~ :
2 . In vi tro: allow ~or adoptive transfer of enriched cell population, especially in - .

- 2 1 2 ~ `) 3 W093/15759 PCT/US93/00~86 regard to G.V.H. reaction.

IV. Decreasin~ Number of SuscePtible Cells ~:-A. Increased levels of ATAA tsee above).

B. Extra-corporeal passage of patient's ~eukocytes over fixed monoclonal ATAA. -:.

C. Use of heterologous antibody directed at :: ~ cellular antigens.

D. Use of pharmacological or biological :
., ~ ~ : entities directed at biochemical or . ..
antigenic constitu~nts of susceptible c~ll. ::

E.: Treat cell suspension in vitro with ATAA to .
~ : . rediuc~the number of susceptible cells.
U ilitY ~ ~:
: 1. Enhance~tumor rejection.
2. Prevention of~tumor development.
The following;~is~a descrlption of the Assay whicb was : used~ iD all experlments detailed in this:application.

DE5CRIPTION~OF ASS~Y ~ :
Essentlally,;~;serum from~an~individual animal is mixed -:~
with a~pool of~ ving~thymocytes from about flve to about :ten::donors. When using inbred animals (Lewis rats), this ~-i5 basically the same ~as:using their own cells since these animals are genetically alike. ~When using:~utbred animal :
; : (Wistar or Sprague~:;Dawley rats) the pool of thymocytes is ~: derived from ab~ut five to about ten individual donoxs all presumably with dlffeFent histocompatibility antigens. The ~ ~ latter was preferred since it mimics the human situation :: :~ ~:

2 ~
WO93/15759 PCT/US93/00~86 where the amount of anti-thymocyte antibody (ATAA) will differ in each individual and it is tested against a common thymocyte antigen present in all individuals within the species. The assay involves mixing a vital dye (trypan blue) with the live thymocytes. The live cells will exclude the dye and will remain unstained (clear) when viewed under a microscope. The dead cells will allow the passage of dye internally so that they ~tain and appear blue. This assay, therefore, reflects the ability of the added serum to kill the target cells. There are innumerable other methods to measure the reaction of antibody in the serum ~anti-thymocyte antibody) with the relevant target cellst i.e. (l ) labeling the target cells with radioactive molecules and observing the release of radioactivity as a function of cell death; (2) measuring .
: : bindin~ of antibody with cells such as using an ELISA ::-assayj (3~ measuring binding of antibody wi~h target cells using the consumption ; of secondary serum molecules : ~ :
: (complement), and: so~ forth. The end result is the :-:
measurement~ of the reaction of antibody in serum with target thymocytes.
. .
Thymus glands were obtained from either Lewis, Wistar or Sprague Dawley rats anaesthetized with ether. The cells were teased~into cold Hanks balanced salt solution, washed twice in the same ~olution and resuspen~ed at concentratiQn of :5X106 cells per: ml. in RPMI 1640-media. After warming at 37C, 100 microliters of cell suspension w~s placed in each well of 300 microliter plates to which was added 50 .

~ 1 2 9 ~

microliters of serum at a 1:3 dilution at 37C. All samples were tested in quadruplicate. Plates were incubated in a stirred water bath at 37C for 30 minutes.
Fifty microliters of cell suspension were mixed with 50 microliters of 0.2% trypan blue and the number of dead and live cells were counted in a hemocytometer plate. One hundred cells from each~sample were separately counted.
EAE was induced by described methods ~J. Neurosci.
:: :
Res. 24: ~ 222 230~ ~1989~] and the end point of dise~se development as a~function~of weight gain is described in detail elsewhere [J.~Pharmacal. Exp. Therap., 242: 614-620 (198~7)]. Adjuvant arthritis was induced as described elsewhere in the~ art ~tProc~ Soc. Exp. Biol . r 137: 506 1971)l. The aforementioned methods only are herein incorporated by these~re~erences.
;It~ should~be~noted ~that the assay system used, al~hough accurate,~is simplistic and economically feasible.
Increased~sensl~tlvity~ ~is~ also achieved by empIoying radioactIve ~labeled~ cells~or various types of bindin~
mea~surement ~technlques.~ The most important assay development~technique~is~to i~dentify the target cell type, and~then purify~the relevant antigenic determinant. Once thi~s is ~accomp~lished, the~ target cell tor antigen) suspension will be~so enriched that, instead of identifyin~
a small number of relevant; cells, most or all will be reactive with the ATAA-co~taining serum. Thus, a more accurate quantitatlon of the amount of ATAA is made . In addition, when the relevant antigen is purified, the ' ::

~129~S
W~g3/15759 PCT/US93/00886 probability is higher that it will be found in more easily accessible cells or tissues, rather than having to use ~.. ;
thymus cells. It is possible to quantitate the number of ;~
susceptible cells with which a person is constituted; in ~`
fact, once the antigen is properly identified it is possible to synthesize it. Thus, pure ATAA as a monoclonal :
antibody, and antigen, as a defined entity, determine the classi~ication of individuals as to their immunological :control mechanisms and therapy will involve the alteration of ATAA and other methods as outlined in Tables A and B.
Although much remains that can be done, it is felt ;-:
that this disc~osure shall serve as a benchmark for future ~
e~forts which employ its: concepts and such is readily :
commended to the field consistent with the hereinafter appended claims.

`
: ~:
':;

-

Claims (9)

WHAT IS CLAIMED IS:

1. A method for evaluating the functional viability of an individual's immune system comprising:
determining an amount of an autoantibody and its regulatory relationship to an amount of its target lymphocyte cell subpopulation; and comparing said amounts to a standard wherein said comparing correlates a high autoantibody amount with a high risk factor for autoimmune disease, vaccine reactions or hypersensitivity reactions and a low risk factor or vulnerability to malignancies, while a low autoantibody amount indicates the probability of high target cell subpopulation, high vulnerability to malignancy and a low cellular immunity.

2. A method for preventing tumor development and/or enhancing tumor rejection by a body comprising increasing autoantibody amount relative to a target cell subpopulation, the target cell being that associated with progression of said development and/or rejection, the increasing accomplished by any of the following:
(a) passively transferring serum from a donor with high levels of said autoantibody to a patient donee (b) producing and transferring to a donee monoclonal autoantibody; and (c) using in the donee immuno stimulants.

3. A method for preventing and/or treating autoimmune disease, vaccine reaction and hypersensitivity reaction comprising decreasing autoantibody amount relative to a target cell subpopulation in a patient/donee, said subpopulation being that of a cell causally related to said disease or said reactions, said autoantibody decreasing effected by one or more of the following:
(a) passing a patient's serum, i.e., extra-corporeal, over antigen containing immuno-absorbents;
(b) administering in vivo a purified target cell antigen; and (c) administering to said patient pharmacological agents which have a function of lowering existing antibody levels.

4. The method of Claim 2 wherein said autoantibody is an ATAA.

5. The method of Claim 3 wherein said autoantibody is an ATAA.

6. The method of Claim 2 wherein said increasing is accomplished by decreasing subpopulation of cells which are ATAA susceptible cells and said decreasing is accomplished by one or more of the following:
(a) increasing levels of ATAA;
(b) extra-corporeal passing of a patient's leukocytes over fixed monoclonal ATAA;

(c) using heterologous antibody by directing same at cellular antigens;
(d) using pharmacological or biological entities which are directed at biochemical or antigenic constituents of said susceptible cell; and (e) treating a cell suspension in vitro with ATAA thereby reducing the number of susceptible cells.

7. The method of Claim 3 wherein said decreasing is accomplished by increasing a subpopulation of cells which are ATAA susceptible cells and wherein said increasing of the susceptible cells is accomplished by one or more of the following:
(a) lowering levels of ATAA;
(b) adoptively transferring said susceptible cells from a fetal or adult source; and (c) deleting, in vitro, other lymphoid cell types thereby causing a functional increase in said susceptible cell subpopulation.

8. A quantitation method for determining the amount of anti T-cell autoantibody, ATAA, relative to a subpopulation of target T-cells in an individual's serum comprising:
(a) obtaining a quantum of target T-cell antigen from an ATAA-recognized antigen pool;
(b) mixing with said quantum of target T-cell antigen a predetermined quantity of the individual's serum;
and (c) determining by conventional quantification methods said amount of anti T-cell autoantibody and the number of target T-cells.

9. The method of Claim 8 further comprising predicting said individual's susceptibility/resistivity to disease or malignancy as such are related to amount of the target T-cells which may be helper, killer, suppressor or other cells involved with immunity in the individual.