Solitary extramedullary plasmacytoma of the palpebral conjunctiva
Factor XIII Deficiency in BALB/c Mice with Plasmacytoma' · American Optical fluorescence...
6
[CANCER RESEARCH 37, 3551-3555, October 1977] 57 ml of plasma obtained from 100 BALB/c mice by a method described by Lowey et a!. (18) for the isolation of human Factor XIII. Factor XIIl-free fibninogen was isolated from human plasma by DEAE chromatography as outlined by Finlayson (10). Purified MFXIII rendered human fibnin clots, obtained by addition of thrombin to Factor Xlll-free fibninogen, insoluble in 5 M urea. Its activity was expressed in units as defined by Lowey et a! (18). Preparation of Antibody to MFXIII. Isolated MFXIII incor ponated into Freund's adjuvant was injected into rabbits at biweekly intervals. Serum of the immunized rabbits was absorbed with mouse plasma free of MFXIII and analyzed for specificity by immunodiffusion. Rabbit antibody to MFXIII caused fibnin clots from human and mouse plasma to be soluble in 5 M urea. Mouse Plasmacytomas. Plasmacytomas MOPC-300, MOPC-384, MOPC-467, and J-558, obtained from Dr. Michael Potter, NIH, Bethesda, Md. , were each trans planted into groups of 30 BALB/c mice. In addition, carci noma line 26, a colon tumor, obtained from Dr. T. Corbett, Southern Research Institute, Birmingham, Ala. , and a spontaneous reticulum cell sarcoma developed in our mouse colony were also used in these studies. At weekly intervals after transplantation 10 mice from each group werebledonce, and theblood wascollected in3.8% so dium citrate (10:1, v/v). Plasma was obtained from groups of mice representing 1-, 2-, and 3-week samples. In addi tion, 10 normal BALB/c mice were bled each week, and their plasma was used as a control for the estimation of MFXIII levels. After 3 weeks, tumor-bearing mice received i_p. injections of 2 mg of cyclophosphamide. The tumors decreased in size and disappeared. These mice were bled 5 and 30 days after treatment with cyclophosphamide. Estimation of MFXIII Plasma Levels. Radial immunodif fusion assays were performed as follows. Antibody to MFXIII was incorporated intoagarose, and10pioftheplasma to be tested were placed into wells 2.5 mm in diameter. Dilu lions representing 100, 50, and 25% MFXIII activity were made from a pool of plasma collected from 20 normal BALB/c mice. The immunodiffusion plates were incubated at 37°for 24 hn, and the diameter of the precipitate was measured. The plasma level of MFXIII was estimated from the linear plot of precipitin diameters obtained with dilu tions of normal BALB/c plasma and reported as percentage of normal. MFXIII activity was estimated in plasma samples of normal BALB/c mice and mice with plasmacytoma ac cording to the method of Coopland et a!. (7), utilizing as well Park Memorial Institute Medium 1640; PBS, phosphate-buffered 55- line (FTA, hemagglutinstion buffer; BBL, Cockeysville, Md.). OCTOBER1977 3551 Factor XIII Deficiency in BALB/c Mice with Plasmacytoma' Joseph Eipe, Vincent Yakulls, and Nicolas Costea Hematology-OncologySection,Departmentof Medicine,UCLASanFernandoMedicalProgram,veteransAdministrationHospital,Sepulveda, California91343 SUMMARY These studies examined the fate of Factor XIII (fibnin stabilizing factor) in mice with plasmacytoma (MOPC-300, MOPC-384, MOPC-467, and J-558). Plasma Factor XIII levels in these mice decreased progressively with tumor expan sion. No plasma inhibitors of Factor XIII activity could be detected . Factor XIII was found on plasmacytoma cell mem branes and in the cytoplasm of the malignant cells by immu nofluorescence. The inverse relationship between tumor load and plasma Factor XIII levels suggested that the fibnin stabilizing factor was absorbed by the malignant cells. INTRODUCTION Factor XIII (fibnin-stabibizing factor), a plasma coagulation factor, when activated by thrombin converts loosely cross linked fibrin into a gel that is insoluble in urea or monochbo roacetic acid and is resistant to the action of proteolytic enzymes (12, 16, 17). This factor plays an important role in wound healing (1) and in the polymerization of collagen (21). Several reports have indicated that a stable clot is an important factor in the establishment and expansion of tumor metastasis (4, 5, 11, 14). It is possible that fibnin forms complexes with platelets, which enhance tumor cell adherence to the vascular endothelium. It has been shown in recent years that interference with fibnin cross-linking can modify in experimental animals the development of tumors and their metastasis (25). In humans, acquired Fac ton XIII deficiency accompanies a variety of malignant tu mors including multiple myeboma (20). It has been sug gested that deficiency of Factor XIII activity in multiple myeboma may result from impaired production or from the presence of antibodies or paraproteins which interfere with its function (20). The results of experiments designed to determine the fate of Factor XIII in BALB/c mice during the growth of trans planted plasmacytoma are herein reported . We present evi dence that plasma Factor XIII decreases in these mice pro gressively with the growth of plasmacytoma and that the malignant cells appear to absorb plasma Factor XIII. MATERIALS AND METHODS Isolation and Assay of MFXIfl2 MFXIII was isolated from I This research was supported by Veterans Administration Research Funds Grant 1577-03. 2 The abbreviations used are: MF)(lll, mouse Factor @IIl@ RPMI 1640, Roe Received March 3, 1977; accepted July 6, 1977. on June 7, 2020. © 1977 American Association for Cancer Research. cancerres.aacrjournals.org Downloaded from
Transcript of Factor XIII Deficiency in BALB/c Mice with Plasmacytoma' · American Optical fluorescence...
[CANCER RESEARCH 37, 3551-3555, October 1977]
57 ml of plasma obtained from 100 BALB/c mice by amethod described by Lowey et a!. (18) for the isolation ofhuman Factor XIII. Factor XIIl-free fibninogen was isolatedfrom human plasma by DEAE chromatography as outlinedby Finlayson (10). Purified MFXIII rendered human fibninclots, obtained by addition of thrombin to Factor Xlll-freefibninogen, insoluble in 5 M urea. Its activity was expressedin units as defined by Lowey et a! (18).
Preparation of Antibody to MFXIII. Isolated MFXIII incorponated into Freund's adjuvant was injected into rabbits atbiweekly intervals. Serum of the immunized rabbits wasabsorbed with mouse plasma free of MFXIII and analyzedfor specificity by immunodiffusion. Rabbit antibody toMFXIII caused fibnin clots from human and mouse plasma tobe soluble in 5 M urea.
Mouse Plasmacytomas. Plasmacytomas MOPC-300,MOPC-384, MOPC-467, and J-558, obtained from Dr.Michael Potter, NIH, Bethesda, Md. , were each transplanted into groups of 30 BALB/c mice. In addition, carcinoma line 26, a colon tumor, obtained from Dr. T. Corbett,Southern Research Institute, Birmingham, Ala. , and aspontaneous reticulum cell sarcoma developed in ourmouse colony were also used in these studies. At weeklyintervals after transplantation 10 mice from each groupwerebledonce,andthebloodwascollectedin3.8%sodium citrate (10:1, v/v). Plasma was obtained from groupsof mice representing 1-, 2-, and 3-week samples. In addition, 10 normal BALB/c mice were bled each week, andtheir plasma was used as a control for the estimation ofMFXIII levels. After 3 weeks, tumor-bearing mice receivedi_p. injections of 2 mg of cyclophosphamide. The tumorsdecreased in size and disappeared. These mice were bled5 and 30 days after treatment with cyclophosphamide.
Estimation of MFXIII Plasma Levels. Radial immunodiffusion assays were performed as follows. Antibody to MFXIIIwasincorporatedintoagarose,and10pioftheplasmatobe tested were placed into wells 2.5 mm in diameter. Dilulions representing 100, 50, and 25% MFXIII activity weremade from a pool of plasma collected from 20 normalBALB/c mice. The immunodiffusion plates were incubatedat 37°for 24 hn, and the diameter of the precipitate wasmeasured. The plasma level of MFXIII was estimated fromthe linear plot of precipitin diameters obtained with dilutions of normal BALB/c plasma and reported as percentageof normal. MFXIII activity was estimated in plasma samplesof normal BALB/c mice and mice with plasmacytoma according to the method of Coopland et a!. (7), utilizing as
well Park Memorial Institute Medium 1640; PBS, phosphate-buffered 55-line (FTA, hemagglutinstion buffer; BBL, Cockeysville, Md.).
OCTOBER1977 3551
Factor XIII Deficiency in BALB/c Mice with Plasmacytoma'
Joseph Eipe, Vincent Yakulls, and Nicolas Costea
Hematology-OncologySection,Departmentof Medicine,UCLASanFernandoMedicalProgram,veteransAdministrationHospital,Sepulveda,California91343
SUMMARY
These studies examined the fate of Factor XIII (fibninstabilizing factor) in mice with plasmacytoma (MOPC-300,MOPC-384, MOPC-467, and J-558). Plasma Factor XIII levelsin these mice decreased progressively with tumor expansion. No plasma inhibitors of Factor XIII activity could bedetected . Factor XIII was found on plasmacytoma cell membranes and in the cytoplasm of the malignant cells by immunofluorescence. The inverse relationship between tumorload and plasma Factor XIII levels suggested that the fibninstabilizing factor was absorbed by the malignant cells.
INTRODUCTION
Factor XIII (fibnin-stabibizing factor), a plasma coagulationfactor, when activated by thrombin converts loosely crosslinked fibrin into a gel that is insoluble in urea or monochboroacetic acid and is resistant to the action of proteolyticenzymes (12, 16, 17). This factor plays an important role inwound healing (1) and in the polymerization of collagen(21). Several reports have indicated that a stable clot is animportant factor in the establishment and expansion oftumor metastasis (4, 5, 11, 14). It is possible that fibninforms complexes with platelets, which enhance tumor celladherence to the vascular endothelium. It has been shownin recent years that interference with fibnin cross-linkingcan modify in experimental animals the development oftumors and their metastasis (25). In humans, acquired Facton XIII deficiency accompanies a variety of malignant tumors including multiple myeboma (20). It has been suggested that deficiency of Factor XIII activity in multiplemyeboma may result from impaired production or from thepresence of antibodies or paraproteins which interfere withits function (20).
The results of experiments designed to determine the fateof Factor XIII in BALB/c mice during the growth of transplanted plasmacytoma are herein reported . We present evidence that plasma Factor XIII decreases in these mice progressively with the growth of plasmacytoma and that themalignant cells appear to absorb plasma Factor XIII.
MATERIALS AND METHODS
Isolation and Assay of MFXIfl2 MFXIII was isolated from
I This research was supported by Veterans Administration Research
Funds Grant 1577-03.2 The abbreviations used are: MF)(lll, mouse Factor @IIl@ RPMI 1640, Roe
Received March 3, 1977; accepted July 6, 1977.
on June 7, 2020. © 1977 American Association for Cancer Research. cancerres.aacrjournals.org Downloaded from
J. Eipe et a!.
substrate human Factor Xlll-fnee fibninogen. The purity ofthe MFXIII preparation was ascertained by polyacrylamidegel electrophoresis (5% gel in 0.03 MTris-barbital buffer, pH8.8). The molecular size of MFXIII was estimated by electrophoresis on sodium dodecyl sulfate-polyacrylamide gel(5.6%) in Tnis-sodium acetate-EDTA buffer, pH 7.4 (9), byusing as standards myoglobin, bovine serum albumin, human IgG, and fibrinogen (24). MFXIII protein isolated frompooled plasma of 100 mice was quantified by the method ofLowry et a!. (19). MFXIII activity of extracts from plasmacytoma was also estimated by the method of Lowey et a!. (18)with fibninogen free of MFXIII as substrate. The tumor extract was obtained by pressing tumor fragments throughwire screens and was collected in RPMI 1640. Tumor cellswerewashed5timeswithPBSandresuspendedin4partscold 0.25 M sucrose. The cells were disrupted in a WaningBlendor, and the supernatant obtained by centnifugation(18,000 rpm at 4°)was dialyzed against PBS and examinedfor MFXIII activity.
The possibility that plasma from BALB/c mice with plasmacytoma contains substances that may inhibit the activityof MFXIII was investigated as follows. Equal volumes ofnormal BALB/c mouse plasma were incubated for 1 h at 37°with equal volumes of plasma from tumor-bearing mice orwith the myeboma proteins that had been isolated by acombination of agar-gel electrophoresis and Sephadex G200 filtration. These mixtures were assayed for MFXIII achyity.Protamine sulfateprecipitation,detectionoffibnindegna
dation products, and platelet counts were done utilizingstandard methods.
Immunofluorescent Studies. Tumors were excised andplaced in RPMI164O, and individual cells were obtained byteasing tumor fragments with scissors. The cell suspensionwas filtered through gauze, placed onto Ficoll-Isopaquesolution, and centrifuged at 2000 rpm for 10 mm at 25°.Thecells on top of the Ficoll-Isopaque layer were removed andwashed 3 times with RPMI 1640. Aliquots of these cellswere dispersed into test tubes to which rabbit antiserumto MFXIII (1:5 dilution) was added. Mixtures were incubated at 25°for 30 mm and then washed 3 times in RPMI1640. To the cell button obtained after the 3rd washing wasadded goat antibody to rabbit lgG conjugated with fluonescein isothiocyanate (3). The goat antibody to rabbit lgG wasdiluted 1:16 for optimal results. After incubation for 30 mmat 25°the cells were washed 3 times and examined with anAmerican Optical fluorescence microscope having verticalillumination (BG 12, kV 418 exciter, and OG 515 barrierfilters). Photographs of cells were taken with Kodak highspeed EktachromefilmEHB 135 (exposuretime60 to 90sec).
For the study of intracytoplasmic MFXIII distribution,plasmacytoma cells were air dried onto microscope slides.The cells were fixed in ethanol at —20°for 15 mm, washed 3times in PBS, and dried. The slides were first flooded withrabbit antibody to MFXIII (1:5 dilution), washed 3 times withPBS, and then overlaid with fluorescent goat antibody torabbit lgG. The slides were washed 3 times with PBS, overlaid with phosphate-buffered glycerol (pH 7), covered, andexamined. Appropriate controls were included, i.e., plas
macytoma cells incubated in normal rabbit serum or withrabbit antibody to fibninogen overlaid with fluorescent goatantibody to rabbit lgG. In some instances plasmacytomacells were incubated with antibody to MFXIII followed byunconjugated antibody to rabbit lgG, and finally the slideswereoverlaidwithfluorescentantibodytorabbitIgG.Cellsfrom tumors other than plasmacytoma and other tissues(liver, kidney, spleen, and bone marrow) were also treatedas described above for detection of MFXIII.
RESULTS
Plasma (57 ml) obtained from 100 BALB/c mice yielded654 @gof MFXIII. Clot-stabilizing activity was estimated tobe 970 units per mg of the MFXIII preparation. Pobyacrylamide gel electrophoresis of MFXIII revealed a single bandmigrating slightly faster than human FXIII. The estimatedmolecular weight of MFXIII was approximately 230,000,whereas human FXIII had a molecular weight of approximately 300,000. Rabbit antibody to MFXIII after absorptionwith plasma free of MFXblbdeveloped precipitin reactionsonly with isolated MFXIII or mouse plasma but not withplasma free of MFXIII as demonstrated by immunodiffusion(Fig. 1). Plasma MFXIII antigen decreased to 48, 36, and28%, respectively, 7, 14, and 21 days after implantation ofplasmacytoma MOPC-467 (Chart 1). A similar phenomenonwasobservedwithplasmafromBALB/cmicein whichplasmacytoma J-558 had been implanted . The fall in plasmaMFXIII antigen in plasmacytoma-beaning mice was statisticabbysignificant when compared with pretransplantationlevels (p < 0.001).
Plasma MFXIII antigen levels of mice transplanted withplasmacytoma MOPC-384 decreased to 41.2% 7 days afterimplant; however, as the tumor grew there was a slightelevation of the antigen (59.7% after 3 weeks). A progressivedecrease of plasma MFXIII antigen was also observed ingroups of 10 BALB/c mice bearing plasmacytoma MOPC300. Cyclophosphamide was injected i.p. 3 weeks after implantation when the level of MFXIII antigen reached 37% ofthe preimplantation value. Five days after treatment, MFXIIIantigen returned to normal values (91.6%) at a time whenmonocbonal lgG was still present in the serum (Chart 2).Plasma MFXIII activity assayed by fibrin clot stability in 5 Murea decreased progressively in parallel with the level ofMFXIII plasma antigen (Table 1).
MFXIII inhibitors were not detected in the plasma of tumon-bearing mice since mixtures of normal mouse plasmaand plasma from plasmacytoma-beaning mice had normalMFXIII activity. Furthermore, increments of myeloma prohem isolated from the serum by electrophoresis and Sephadex G-200 filtration from mice with tumor when added tonormal mouse plasma did not reduce the activity of MFXIII.
Since the protamine sulfate test and platelet counts werewithin normal limits and the search for fibnin degradationproducts yielded negative results, disseminated intravascuIancoagulation was not present in the tumor-bearing mice.
Plasma cells isolated from different plasmacytomas wereincubated with antibody to MFXIII followed by goat antirabbit lgG conjugated with fluorescein isothiocyanate.
CANCER RESEARCH VOL. 373552
on June 7, 2020. © 1977 American Association for Cancer Research. cancerres.aacrjournals.org Downloaded from
I
Factor XII! in Mouse P!asmacytoma
!:. ‘
-.@. S
r•@5.
!@ ,@‘
even, megakaryocytes and platelets contained MFXIII whentreated in a similar fashion.
Sucrose extracts from plasmacytoma cells containedMFXIII activity as demonstrated by their ability to renderinsoluble in 5 M urea Factor XllI-free fibnin clots. Furthermore, the activity of these extracts was inhibited by priorincubation with antiserum to MFXIII. Cells isolated fromtumors other than plasmacytoma (carcinoma line 26 andreticulum cell sarcoma) did not carry MFXIII. In addition,their plasma MFXIII levels and activity were unchanged duning tumor growth.
DISCUSSION
The results of our experiments indicate that in mice withplasmacytoma the level of plasma MFXIII decreases withexpansion of the tumor. This phenomenon was observed inall mice bearing plasmacytoma regardless of the type ofparaprotein synthesized by the tumors. After eradication ofthe plasmacytoma by i.p. injection of cycbophosphamide,MFXIII plasma levels returned to normal values.
In patients with multiple myeloma, the decreased MFXIIIlevel was considered to be due to the inhibitory effect ofparaprotein which may have interfered with the functions ofthis coagulation factor (22). Results of our experimentsindicate that in mice with plasmacytoma there was no interaction between myeboma proteins and the activity of MFXIII,since both MFXIII activity (as determined by clot stability inSMurea)andantigenlevelsreturnedtonormalvaluesafterthese animals were treated with cycbophosphamide at atime when monocbonal Ig was still present in their sera.Results of mixing experiments in which normal mouseplasma was incubated with monoclonal Ig isolated from the
(10)
100
75
50
25
I
OCTOBER1977 3553
‘MtP'@H‘-xII!@.
HPS I
M FIBQ@@ .
Fig. 1. Immunodiffusion of MFXIII (XIII), mouse plasma (MP), mouse plasma free of MFXIII (MP-Xlll), mouse flbnnogen free of MPXIII (M FIB), and humanplasma (HP) developed by antibody (ab) to MFXIII.
(10)
ft (10)
(10)
0@7 14 21
days after tumor transplantChart 1. Plasma MFXIII levels after implantation (arrow) of MOPC-467 into
BALB/c mice.
Plasma cells from plasmacytomas J-558, MOPC-467, andMOPC-384 demonstrated surface and cytoplasmic staining(Fig. 2). Most of the cells had a faint circumferential fluorescentringwithsuperimposedbrightspots.Plasmacellsofplasmacytoma MOPC-300 exhibited only cytoplasmic fluorescence.In sometumorsnumerousplasmablastscontamed coarse intracytoplasmic fluorescent staining (Fig. 2).Plasmablasts incubated with normal rabbit serum or rabbitantibody to fibrinogen before the addition of fluorescentgoat antibody to rabbit IgG did not fluoresce. Incubation ofplasmablasts with antibody to MFXIII followed by addition ofunconjugated goat antibody to rabbit IgG decreased appreciably the immune fluorescence after fluorescent antibodyto rabbitlgGhadbeenadded.
Cell suspensions from liver, spleen, and kidney failed todemonstrate fluorescence with antibody to MFXIII; how
on June 7, 2020. © 1977 American Association for Cancer Research. cancerres.aacrjournals.org Downloaded from
Decline of MFXIII antigen and activity in BALB/c mice (N = 40) withplasmacytomaMOPC-300Days
aftertumortrans Radial immunodiffu
plantsiona Clot stabilityassayb097.4
± 7@9c 94.0 ±12772.3±14.1 76.4 ±15.41450.9± 4.2 48.2 ±12.22137.0± 3.9 32.4 ± 8.4
J. Eipe et a!.
(10)
.1017 14 21@ 26
CTXdaysafter tumor transplant
Chart 2. Plasma MFXIII levels after implantation (arrow) of MOPC-300 inBALB/c mice and after i.p. injection of cyclophosphamide (CTX).
(endocytosis) of this coagulation factor by the malignantplasma cells. It is also conceivable that MFXIII might havebeen incorporated as part of the transamidation of polymerized fibrin deposited during the establishment of the tumon. We have not detected MFXIII in plasma cells of splenicon bone marrow origin of tumor-bearing mice. Clot-formingand -stabilizing activity was also found in a fraction fromplasma cell tumor YPC-1 (13).
Several reports have indicated that deposition of polymenized fibnin plays an important role in the growth ofmalignant tumors and the establishment of their metastasis(4, 6). Studies in both humans and experimental animalsindicate that interference with fibnin deposition netardsgrowth and metastasis (2, 15, 25). Interference with theformation of a fibnin network and stabilization by Factor XIIImay represent a rational approach for the treatment of certam tumors. Factor XIII absorbed by malignant plasma cellsmay also represent a propitious target for active on passiveimmunotherapy.
REFERENCES
1. Beck, E., Duckert, F., and Ernest, M. The Influence of Fibrin StabilizingFactor on Growth of Fibroblasts in Vitro and Wound Healing. Thromb.Diath. Hsemorrh., 6: 485-491 , 1961.
2. Brown, J. M. A Study of the Mechanism by Which Anticoagulation withWarfarin Inhibits Blood-borne Metastasis. Cancer Res., 33: 1217-1224,1973.
3. Cerottini, J. C., and Webb, T. Preparation and Use of Specifically Punfied Antibodiesfor lmmunofluorescentLabelling.Acts Histochem.,27:225-234, 1967.
4. Chew, E. C., and Wallace, A. C. Demonstration of Fibrin in Early Stagesof Experimental Metastasis. Cancer Res., 36: 1904-1909, 1976.
5. Clifton, E. E., and Agostino, D. Factors Affecting the Development ofMetastatic Cancer. Effect of Alterations in Clotting Mechanism. Cancer,15: 276-282, 1962.
6. Clifton, E. E., and Agostino, D. The Effect of Fibrin Formation andAlteration in the Clotting Mechanism on the Development of Metastasis.Vssc. Dis., 2: 43-52, 1965.
7. Coopland, A., Alkjsersig, N., and Fletcher, A. P. Reduction in PlasmaFactor XIII (Fibrin Stabilizing Factor) Concentration during Pregnancy. J.Lab.Clin.Med.,73:144-153,1969.
8. Duckert, F. Documentation of the Plasma Factor XIII Deficiency in Man.Ann. N. Y. Acad. Sci.. 202: 190-199, 1972.
9. Fairbanks, G., Steck, T. L., and Wallach, D. F. Electrophoretic Analysisof the Major Polypeptides of the Human Erythrocyte Membrane. Biochemistry, 10: 2606-2617, 1971.
10. Finlsyson, J. S. Human Fibrinogen Free of Factor XIII: The Pursuit ofBiological Zero., Ann. N. Y. Aced. Sci., 202: 149-154, 1972.
11. Fisher, B., and Fisher, E. R. Experimental Studies of Factors WhichInfluence Hepstlc Metastases. VIII. Effect of Anticosgulants. Surgery, 50:240-247, 1961.
12. Laki, K., and Lorand, L. On the Solubility of Fibrin Clots. Science, 108:280,1940.
13. Laki, K., Tyler, H. M., and Yancey, S. T. Clot Forming and Clot StabilizingEnzymes from the Mouse Tumor YPC-1. Biochem. Biophys. Res. Cornmun., 24: 776-781, 1966.
14. Larsen, V., Mogensen, B., Amris, C. J., and Strom, 0. FibrinolytlcEnzyme in the Treatment of Patients with Cancer. Danish Med. Bull.. 5:137-140, 1961.
15. Larsen, v., Mogensen. B., Amris, C. J., and Strom, 0. FibninolyticEnzyme in the Treatment of Patients with Cancer. Danish Med. Bull. , 5:137-140, 1964.
16. Lorsnd. L. Properties and Significance of the Fibrin Stabilizing Factor(F5F). Thromb. Diath. Hsemorrh., 7 (Suppl. 1): 238-247. 1962.
17. Lorand, L., and Jacobson, A. Studies on the Polymerization of Fibrin.The Role of the Globulin: Fibrin Stabilizing Factor. J. Biol. Chem., 230:421-434, 1958.
18. Lowey, A. G., Dunsthan, K., Kriel, R., and Wolfinger, H. L., Jr. FibrlnssePurification of Substrate and Enzyme. J. Biol. Chem., 236: 2625-2633,1961.
19. Lowry, 0. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. ProteinMeasurement with the Folin Phenol Reagent. J. Blol. Chem., 193: 265-275,1951.
I
Table 1
a MFXIII estimated by radial immunodiftusion (percentage of
normal values).b MFXlll estimated by clot stability assay (percentage of normal
values).C@ S.D.
serum of tumor-bearing mice confirmed the observationthat the paraproteins synthesized by tumors MOPC-300,MOPC-384, MOPC-467, and J-558 did not interfere withMFXIII activity.
The accumulation of MFXIII on on in plasmablasts wasdemonstrated by immunofluorescence, and extracts of allplasmacytomas investigated yielded active MFXIII.
Reduction of plasma Factor XIII has been observed in avariety of conditions (8). In congenital Factor XIII deficiency,although a quantitatively near-normal amount of this factorhas been detected in the plasma, its function was abnormal,probably because of synthesis of a defective molecule (23).In mice with plasmacytoma, synthesis of defective MFXIIImolecules is unlikely since assay for quantitative and qualitative changes revealed a progressive decline of both MFXIIIactivity and antigen.
We have also investigated the possibility that disseminated or localized intravascular coagulation may have beenresponsible for the reduction of this consumable coagulalion factor. We did not detect in the plasma of tumorbearing animals by-products of intravascular coagulation,nor could we demonstrate fibnin on the surface of malignantcells. Deposition of fibnin in and around tumors appears tobe a transient phenomenon (4). We conclude that reductionof MFXIII in mice with plasmacytoma is due to cellularuptake of this factor and in some instances by ingestion
3554 CANCERRESEARCHVOL. 37
on June 7, 2020. © 1977 American Association for Cancer Research. cancerres.aacrjournals.org Downloaded from
Factor XIII in Mouse P!asmacytoma
20. Mandel, E. E., and Gerhold, W. M. Plasma Fibrin-stabilizing Factor:AcquiredDeficiencyin VariousDisorders.Am.J. Clin. Pathol.,52: 547-556, 1970.
21. Nyman, D., and Duckert, F. Factor @lU.Fibrin and Collagen. Thromb.Diath. Haemorrh., 34: 551, 1975.
22. Penny, A., Castaldi, P. A., and Whitsed, H. M. Inflammation and Hsemostasis in Paraproteinsemias. Brit. J. Haematol., 20: 35—44,1971.
23. Sonia, J., Soria, C., and Samama, M. Presence dun AntigénePlasma
tique Indispensable a Is Stabilisation de Is Fibrine Dans Deux Cas deDeficit Congenital en Facteur XIII. Thromb. Disth. Hsemorrh., 24: 356,1970.
24. Weber, K., and Osborn, M. The Reliability of Molecular Weight Determinations by Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis. J. Biol.Chem., 244: 4406-4412, 1969.
25. Yancey, S. T., and Laki, K. Transglutsminsse and Tumor Growth. Ann.N. Y. Aced. Sci., 202: 344-348, 1972.
2A
2CFig. 2. Immunofluorescent staining of tumor cells with rabbit antibody to MFXIII and goat antibody to rabbit IgG labeled with fluorescein isothiocyanate.
Staining of viable cells (A, B) and fixed cells (C, D).
OCTOBER1977 3555
2B
Si
2D'@5@
5fr1P@
on June 7, 2020. © 1977 American Association for Cancer Research. cancerres.aacrjournals.org Downloaded from
1977;37:3551-3555. Cancer Res Joseph Eipe, Vincent Yakulls and Nicolas Costea Factor XIII Deficiency in BALB/c Mice with Plasmacytoma
Updated version
http://cancerres.aacrjournals.org/content/37/10/3551
Access the most recent version of this article at:
E-mail alerts related to this article or journal.Sign up to receive free email-alerts
Subscriptions
Reprints and
To order reprints of this article or to subscribe to the journal, contact the AACR Publications
Permissions
Rightslink site. Click on "Request Permissions" which will take you to the Copyright Clearance Center's (CCC)
.http://cancerres.aacrjournals.org/content/37/10/3551To request permission to re-use all or part of this article, use this link
on June 7, 2020. © 1977 American Association for Cancer Research. cancerres.aacrjournals.org Downloaded from
