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SIMILAIT DES AND PITTARKHIS IN TIX PECITIC IMMINOLVICAL
SREDOV: IVENESS ANTE
NETICALLY TOLERANT UCE AND MIQSA MDE TOLERANT AT BIRTIR
Jan Wright cairan
Hion 3 Divisix, Qui Ricije? Yae! amil Libor.tur: Quk Ricst, 18:nesia
LEGAL NOTICE
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Research spansored by the U. S. Atanic Berøy Cunnission under cuntract with
tbe Union Carbide Corporation.
The unresponsiveness of an I, hybrid jouse to skin grasto of
either parental type can be allod natural or genetic talerance, and
uther states of unresponsivenesa, including induced, o: actively-ucquired,
tolerance rawy profitably be capared to it. Time posolbility that in
antigen or antigens exist in i parental strain that fail to be expreciou
In an P, lwybrid is an interesting one, but it is in direct caiflict with
the abservation or canetic tolerance to skin crafts. Thio possibility of
parant-specific witigens 'de's boon invokord to explain experimental results
not directly predictable fru. the so-callid lave of transplantaiian
(Celad:r and Walshans, 1942; Cudkowicz, 1961, 1964).
The folloding experiments were designed :o search for purental
antigas not expressed in Flybride and to compare responsive_jess of
tolerant parental mice with that of genetically tolerant , hyhrids.
Results concerning induction by my spleen cells of tolerance to parental
skin, together with data an serum antibody production, have been
published elsewhere (submitted for publication, coatiman). The results
presented here deal primarily with the histology of wresponsive or
tolerant mica that have been given spleen injections.
METHODS
Parental otruin mice, within 12 hours of birth, were dich givan
intraperitoneally approximately 20 million splavan calls in .05 m?
saline from I, hybrid malos.
Some of the nice, when they were 3-9 months ald, were given rull
thickness car granto on the dorsal portions of the lateral thoracic walls.
Others, presumably-tolerant, were not given skin grafts.
Mouse strains and Hyorlds usod included C57BL/6 Cum (cullod Blo),
DB/2 cm (2), (B) + x 2 + (BOT): c57BL Ca (B), c3N/An cưa (23),
18 + xc3 os)?, (BC #P), A.CA, aró A.31.
jerrizm ves obtained from tail artery or heart blood trust had boun
slovod to clot at room temperature. Hangasglutinations and cytotoxic
activities wore determired by the stadard nothioco (vorer and Mikulaka,
19543 Marer and O'Dortuuli, 1950).
wplec. coll juopancions were propard by prusoins the organ through
a screen and then dispersine mall cliems by gentle topiration with a
syringe through a 25-gauge needla. Cells were counted in a ham aytameter
and were appropriately dliuted for intraperitoneal injectim in a volume
of 1 milu liter per adult puude.
Individual paly weights vore recorded two or thron times a week in
experiments in which wasting diocase might be anticipated.
Mice were decapitated before necropoy. Tissues were fixod in
zankor-formol solution, and sections were stained with Sonataxylin and
dain.
RESULTS AND DISCUBSION
The graft-varsus-bost diacuse that nogults from the injection or
parental oploen cells into I, hybrids has been described several timeo
(Doogerove ot al., 1968; Barer and Boyse, 1959; Kaplan and Rooston, 1959;
Plocus et al., 1966). ja ymeral, the same findings were made ia the
wasting disease arter receiving 20% oploce collo from either parental strain.
The reason for this failure 18 not understood, but the finding 18 not
altogether unexpected, inasmuch as variability in incidence and coverity
of secondary dlugabe among different para..), jwybrid combinations has been
reported. The other lybrid studied here, Beiber, cid undergo wusting
diocase aster receiving purental Ef...en cellu. Body weight be. ....."
began within the second week aftor injection and amounted to 20-40% or
the initial valucz by the t.lme nice were found doun or woru kilied for
further study, usually 3 to 5 weeks after receiving spleen. Anenia
accompanied the weight loss, end hematocrito at the time of death were
in the range of 20.37%. Moreover, heaaeglutination studies with specific
anti-parent sora on these animals indicated that many of the red collo
vore derivad from the donor spleen coll inocule. Consistent with this
anemia, marrow was either hypoplastic wr completely aplastic (P18. La).
Severe atrophy occurred in the lymph nodes and spleen white pulp of
recipients of parentai spleun (Fig. lb, c).
To erant or presumably-tolerant paren, 1 mice, after receiving second
parental spieen colls, did not all behave o imilarly. Many appeared to
dovelop wasting disease, but others showed no weight loss at all. Some
of the inaryocted ones, presumably-tolerant but not skin grattad, may not
have been tolerant at all. But 13 of the group not showing wasting disease
carried second parental car grafts. The inconsistency of these resuits 18
perplexing. It is, of course, possible that despite persistence of
foreign skin grafts, a degree of histoincompatibility existed permitting
rejection by the host of donor spleen cells. However, factors that are
not inunnalogical muy isloo influence tiw tuirsicturce of colls, and, 11
tive abocnce of definitive evidmoo, it would thorcroru ucom 111 -udviwari
to uscribe the failure to an lumologic seclusion at thio time.
Thore tolorunt juarenta) nice that did devi:lop wuotinus dineuse
bugun to lose weight within two to thate wecke 1..or receivinas oscand
parental splorn, and they praygrossively Luot iu a wivy comparable to
similarly trusted P, kidults. 'irouo nico aluo vecuma soverely anamic
bofora doath. However, thro were soveral striking histological different
noted between the tiro grupo. intherous narrov spucos of P, 's were usually
empty or nearly 30, those of tolerant mico word only slightly hypocollular
and være regularly 'yperkrinulocytic (F18. 24). Parental nice, although
they had doplotou lyzwh nodes and spioon white pulp, as a rulo did not show
Buch extremo lyphoid urophy as did analogous ? hybride. Occasionally
some regeneration could be l'ound In lymphatic trasue, and gorum cantero,
although not cannon, were sometimes sot (818. 20). Agyloidosis, often
soon after a chronic infection or other proionged from inte atimulatia,
was a frequent finding in sploons (P18. 2c) and livers of tolerant parental
nooipients of second parental apleea cells but not in tissues ut !, recipients.
in general, parental Ace witiba wasting discaso save the impression of
having undergone a more protracted Inume response, presuably graft-voraus-/1087,
than rad P, hybride'. Hybrids appeared to have ouflared an acute, ueruotating
attenk from which no recovery was possible. Tolerant parental noe, either
were subjected to a look severe attack, poweibly by virtue of offering a
loss hospitable wvironment to the oploma cell grant, or were able to
rocover to form extorit fran the dumpir inflicted.
One theoretical possibility CC9108 to mind to demont for the
histological differances Boon between genoticully and inductively tolorant
mice. Perhaps second parontal oploon cells, toloratod by the othor parental
strain at "irat, in tho proceso of grutt-verouo-liunt reuction doutroy the
progony of I, hybrid cells that were injectyd ut birth. With the dotruction
of these cells, tolerance would be broken, and the host could then reject
the donor spleen colls and their descendenta. No stronger cubo cv de w
e
for the probability of this lvypothesis than to gint out that it is:
consistent with the histologic findings and vitli the lack of erythrocyte
chimerion among tolerant parental mice.
SUMMARY
In the course of a search for parental anticeno not expressed in P,
hybrid mice, observations were made on two kinds of wresponsiveness:
genetic, or that of a hybrid toward ito parents, and induced or actively-
acquired, exemplified here by a parental strain made tolerant to a second
parental strain by receiving bybrid cells at birth. Both groups of nice
failed to pruduce circulating bemagglutinins to second parental cella, and
this failure was specific. Mice from both groups inderwent wasting disease
when given an injection as adults or second parental aploen cells.
Histologically, lymphatic tissues of tolerant parental mice appeared less
Beverely atrophic than those of genetically tolerant P,'s. Their marrow
was regularly more cellular than that of hybrida sirdlarly treated. A
theoretical possibility, involving the breaking of tolazance in parental mice,
18 offered to explain the abaorved differences been between two groups that
szight have been expected to behave similarly.
ACKNOWLEDIMENT
The author 18 grateful to Dr. C. C Congdon for invaluable
help in evaluating and photographing histological material.
RETERXNCES
1. Colada, Franco, and Welshans, W. J. 1962. Proc. Nat. Acad. Sci.
48: 326.
2. Cudkowicz, Gustavo. 1961. Proc. Soc. Exp. Biol. Med. 107: 968.
3. Cudkowicz, Guctavo. 1964. Fed. Proc. 23: 202.
4. Goodman, Joan Wright. A search for parental antigens not expressed in ?
hybrid mice. (Submittes for publication).
5. Gorer, P. A., and Mikulska, 2. B. 1954, Car.cer Research 14: 651.
6. Gorer, P. A., and O'Gorman, P. 1956. Transpl. Bull. 3: 142.
7. Cosgrove, G. E., Upton, A. C., and Congdon, C. C. 1962. A. J.
Pathol. 40: 455.
8. Gorer, P. A., and Boyse, E. A. 1959. Immunol. 2: 182.
9. Kaplan, Henry S., and Roseton, Barbara H. 1959. Stanford Mod. Bull.
17: 77.
20. Piscus, W. G., Morris, B. T., Jr., 348810a, J., and Trentin, J. J.
1962. Ann. N. Y. Acad. Sci. 99: 355.
PICURE LEGENDE
Pig. l. Sections from un adult B6D2r, hybrid male 17 days after receiving
10° B6 sploen cells intraperitoneally.
4. aplastic femoral marrow space. X. 290.
b. atrophic lymph node. X 290.
6. severely depleted spleen. X 115.
Fig. 2. Sections from three different tolerunt parental mice tú veeks
after receiving 100 second parental spleen csils intraperitonealiy.
a. cellular, hypergranulocytic fcrcrul nrrow. X 290.
b. depleted Lymph node with glicna cells and area of regeneration.
X 290
6. Extreme amyloidosis of spleen. X 115.
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DATE FILMED
11/ 20 / 64





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END