[CANCER RESEARCH 46, 271-277, January 1986]

Quantitative Whole-Body Autoradiography of Radiolabeled AntibodyDistribution in a Xenografted Human Cancer Model1

Irwin Fand, Robert M. Sharkey, William P. McNally, A. Bertrand Brill, Prantika Som, Kazutaka Yamamoto,F. James Primus, and David M. Goldenberg2

Department of Psychiatry and Behavioral Science, State University of New York, Stony Brook, New York 11794 [l. F., W. P. M.]; Institute of Molecular Immunology, Centerfor Molecular Medicine and Immunology, University of Medicine and Dentistry of New Jersey, Newark, New Jersey 07103 [R. M. S., F. J. P., D. M. G.¡;and MedicalDepartment, Brookhaven National Laboratory, Upton, New York 11973 [A. B. B., P. S., K. Y.¡

ABSTRACT

Quantitative whole-body autoradiography (WBAR) was usedto study the biodistribution of goat anti-carcinoembryonic antigenand normal goat IgG, each labeled with 125I,in hamsters bearing

the carcinoembryonic antigen-producing GW-39 human coloniecarcinoma xenograft. Comparisons between computer-assisted

videodensitometric profiles of WBARs and tissue radioactivitycounts were made at 1, 3, and 7 days following administrationof the radiolabeled IgGs. The results indicated that maximaltumor accretion of the radiolabeled antibody and normal IgGoccurred within 1-3 days, with a marked selective accretion ofantibody in the tumor being evidenced at 3-7 days because of

clearance of normal IgG. Radioactivity derived from antibody IgGshowed 6.5 to 118.7 times that found in other tissues, asmeasured by videodensitometry, whereas organ radioactivitycounting revealed ratios of only 6.7 to 29.6. Specificity of tumor-

cell accretion of the radiolabeled antibody was confirmed bymicroscopic autoradiography, showing intense labeling of theproliferating perimeters of GW-39 tumors. WBAR was found tohave a resolution of 0.10 to 0.25 mm in 100-g hamsters, which

appears to be greater than the resolving power of external bodyimaging by gamma camera scintigraphy. These studies suggestthe use of WBAR and microautoradiography to complementexternal imaging methods for the analysis of antibody distributionand localization in cancer radioimmunodetection models.

INTRODUCTION

Previous studies of cancer localization with polyclonal andmonoclonal antibodies have used tissue counting (1-6), microautoradiography (7-9), and external body imaging (1, 2, 10, 11)

techniques. External immunoscintigraphy in humans has received considerable attention because of the demonstration thatcancer sites can be disclosed after the administration of radio-

labeled polyclonal and monoclonal antibodies directed againstcancer-related antigens (12-14). Indeed, there is evidencemounting that external immunoscintigraphy, also termed RAID,3

(15,16), may be the method of choice for detecting occult cancersites (12, 13, M-22). However, even animal studies of cancer

imaging do not provide sufficient information on the cellular and

Received 6/3/85; revised 9/30/85; accepted 10/4/85.The costs of publication of this article were defrayed in part by the payment of

page charges. This article must therefore be hereby marked advertisement inaccordance with 18 U.S.C. Section 1734 solely to indicate this fact.

1Supported in part by NIH grants CA37412 and CA37849.2To whom requests for reprints should be addressed, at the Center for Molecular

Medicine and Immunology, 100 Bergen Street, Newark, NJ 07103.'The abbreviations used are: CEA, carcinoembryonic antigen; RAID, radioim

munodetection; WBAR, whole-body autoradiography.

tissular distribution of the antibody preparations. In contrast tothe ¡magesprovided by gamma camera photoscans, WBARpreparations allow an appreciation and comparison of the topographic and tissue distribution of radioactivity in animals receivingradiolabeled antibody and control immunoglobulin preparations,thus permitting a better understanding of the non-uniform deposition of radioactivity within a tumor or organ. Since the CEA-producing GW-39 human colonie carcinoma system has formedthe basis of most of our RAID studies with anti-CEA IgG (2, 5,

6), it was of interest to investigate the application of qualitativeand quantitative WBAR in this model system. The results indicatethat this method provides a better understanding of the basis oftumor accretion of radiolabeled ¡mmunoglobulins.

MATERIALS AND METHODS

Tumor System and Radioantibody. Female golden hamsters (Mesocricetus auratus) weighing 70-120 g were inoculated with 0.5 ml of a10-20% cell suspension of CEA-producing GW-39 human colonie car

cinoma cells (23, 24) s.c. in the nape of the neck. After 6 days, pairs ofanimals were given injections i.p. of 60 ¿iCiof 125l-goat anti-CEA or 125I-

normal goat IgG. Goat anti-CEA IgG was purified by affinity chromatog-

raphy as described previously (5, 13), whereas normal goat IgG waspurchased from Miles Laboratories (Elkhart, IN) and used without furtherpurification. Both IgG preparations were labeled with 125I(Amersham,

Arlington Hts., IL) by the chloramine-T procedure (25) to a specific activityof 12 Ci/g. After removal of unreacted iodine by gel filtration over a PD-

10 column (Pharmacia), approximately 60% of the radioiodinated antibody bound to a CEA immunoadsorbent, while only 5% of the normalgoat IgG bound to the adsorbent. Greater than 90% of the radioactivityfrom both preparations bound to an anti-goat IgG immunoadsorbentand, by gel filtration on S-200 (Pharmacia), greater than 95% of each

preparation migrated as native IgG.Whole-Body Autoradiography. After pentobarbital anesthesia, the

tumor-bearing hamsters were flash-frozen in liquid nitrogen and em

bedded in carboxymethylcellulose. After trimming to the midline regionin the sagittal plane, 50-Mm sections, at depths of approximately 250^m, were made with an LKB-2258 cryomicrotome at -17°C. The sec

tions were mounted on transparent tape, freeze-dried for 3-4 days at-17°C, and then placed directly on LKB Ultrofilm for 8 days at -20°C.Exposure was made for 1-2 weeks at -17°C without the use of

intensifying screens. Completed macroautoradiograms were photographically processed using standard film developing methods. Theseprocedures have been reported earlier (26, 27).

Microautoradiography and Tissue Counting. Separate groups ofanimals were used for microscopic analysis of radioactivity grains. Atdays 1,3, and 7 following radioantibody administration, the animals weresacrificed, and the tumors and organs were removed for counting. Thetumors were placed directly in a scintillation vial containing 10% bufferedformalin. After counting the 125l-radioactivity, the tissues were embedded

in paraffin, and 5-^m sections were cut and mounted on glass slides by

conventional procedures. The slides were then processed for microautoradiography according to methods published previously (28, 29).

CANCER RESEARCH VOL. 46 JANUARY 1986

271

Research. on August 30, 2021. © 1986 American Association for Cancercancerres.aacrjournals.org Downloaded from

RADIOANTIBODY WHOLE-BODY AUTORADIOGRAPHY

Briefly, deparaffinized sections were hydrated by passage through serialdilutions of ethanol. The slides were then dipped into an aqueous (1:1)dilution of Kodak NTB-2 emulsion in the darkroom. The dipped slides

were then dried for 45 min in a cool stream of circulating air with relativehumidity maintained at 75-90%. Exposure was made in light-tight boxesfor 2-3 weeks. After exposure, the slides were processed for photography using D-19 developer and GBX fixer at 15-19°C. The slides werethen post-stained with Harris' hematoxylin-eosin. A series of slides

without sections, but otherwise treated identically, were included toassess possible chemography artifacts.

Videodensitometry. A Hamamatsu video camera interfaced with aPOP 11/34 computer equipped with GAMMA-11 display system softwarewas used for quantitation of the autoradiograms. By digitizing a step-

wedge series of autoradiographed radioisotopic standards, a responsecurve relating the level of activity with the digitized Hamamatsu number(an arbitrary number produced by the video camera) was constructed.The computer used this information to calculate the actual amount ofradioactivity in each pixel of the digitized matrix. Digital sampling waspossible from 256 x 256 to 1024 x 1024 with a 256 gray scale. Thefinal step in calculating the levels of activity in each pixel of the digitizedimage involved performing a polynomial fitting to the standard calibrationcurve or by linear interpolation. The quantified image was then displayedby color coding 16 different levels of activity. Regions of interest werethen selected by using the GAMMA-11 system to analyze the autoradi

ograms. This permitted a high spatial resolution of the photographedimage in the digitalized image. Ten or more pixels were sampled,digitized, and converted to mCi using an autoradiographic standardcurve. From these values, the means and standard derivatives of thesamples were calculated. Blood pool activities were determined in cry-

osections in cardiac cavities and/or abdominal aorta.For preparation of the autoradiographic standards, an arithmetic dilu

tion series of 16 intervals was prepared for 125I,using a modification of

the method of Cross ef al. (30). Briefly, strips of Kodak SB-5 filmpreviously fixed without hardener were soaked for 5-15 min in the

serially diluted aqueous solutions containing radioiodine, were driedovernight, and were aligned together to form standard strips. Thesestrips were then placed against X-ray film simultaneously with cryosec-

tions from the animals and exposed and processed simultaneously. Toobtain dpm/mm values of the step-wedge 125Idilution series, uniform

punches were taken from the standard strips, and the radioactivity wasdetermined in a well-type gamma-scintillation counter.

RESULTS

WBAR at 1 day post-injection of the radiolabeled preparations

does not show any distinction between tumor uptake of antibodyor normal IgG, but the uptake of the radioactivity in both preparations is clearly higher in the tumors than in other organs of thehamsters (Fig. 1). On the third day, however, a differential uptakein tumor between the antibody and control IgG is beginning tobe appreciated (Fig. 2). By 7 days, the radioiodinated anti-CEA

antibody is retained selectively in the tumor, while only a weakcentral zone of activity is seen with the normal IgG (Fig. 3). Theareas of highest radioactivity obtained with normal goat IgG areseen in the tumor's central, cystic structures (Fig. 3, b and d).

By measuring sites of radioactivity in the tumors, it is apparentthat areas as small as 0.10 to 0.25 mm could be demonstratedby this method. Radioactivity observed in the extra-tumor re

gions (liver, heart, muscle, kidney, spleen, etc.) on days 1 and 3was essentially uniformly distributed. At day 7, radioactivity inmost extra-tumor sites was reduced to levels near background.

These WBAR results are corroborated by microscopic auto-

radiography, which shows the location of radioactivity in theperipheral portions of the tumor, associated with viable cells andin the proximity of blood vessels (Fig. 4), whereas the radioactiv

ity seen in the tumors receiving the control IgG is found moreuniformly distributed throughout the tissue sections with appreciably less grain disposition. Control emulsion-coated slides with

out tissue section showed no artifacts of chemography.Videodensitometric quantitation of the WBAR sections re

vealed a similar distribution of the antibody and control IgGs onday 1 (Table 1). Although the concentration of radioactivity washighest in lung, tumor, and blood, tumors could be distinguishedby either the antibody or control IgG at this time (Fig. 1). Examination of the results of day 3 and day 7 sections, however,indicated that specific tumor accretion increased with time withthe anti-CEA antibody preparation, ranging from a mean tumor/non-target ratio of 89.4 for muscle to 9.7 for blood (day 7). In

contrast, the mean tumor/nontarget ratios for the control IgGwere a high of 17.8 for muscle and a low of 2.8 for blood at thissame time, thus providing a specific tumor localization ratio(antibody/control IgG) of 5.0 for muscle and 3.5 for blood.

It became apparent upon analysis of the Videodensitometricresults that radioactivity accretion and, in turn, tumor/non-targetratios could vary considerably, depending on the zone of reference used within a tumor section; i.e., whether areas of leastradioactivity corresponding with sites of diminished viability or ifperipheral tumor cell clusters were chosen. Therefore tumor/non-target ratios were constructed by comparing results from

the lowest densitometric values to those derived from the highestvalues in the tumors (Table 2). These differences can vary about2-fold for specific antibody to even 3-fold (day 7) for the control

IgG.A comparison of the findings of WBAR Videodensitometry and

tissue radioactivity determined by direct counting of specimenaliquots can be seen by evaluating the counting results (Table 3)derived from comparable tissues used for Videodensitometry(Table 2). It is apparent that the tumor/non-target ratios foundby direct counting of cpm/g are consistent with the densitometricresults for the areas of lowest activity in the tissues (Table 2).This may be due to the aliquots of tissues used for direct countinghaving a relatively low abundance of radioactivity, since the latteris more restricted to actively proliferating zones of tumor cellswhich are likely to be less available for tissue sampling. Thelarger the tumors, presumably the smaller the relative amount ofantibody-accreting tumor cells available for analysis. According

to these studies, microscopic autoradiography or quantitativeWBAR provides a more accurate assessment of tissue localization of antibody-transported radioactivity.

DISCUSSION

CEA was the first defined cancer-associated marker to serve

as a target molecule for radiolabeled antibodies in clinical cancerRAID (13, 15). Subsequent studies have confirmed the usefulness of CEA antibodies for RAID (12, 14, 18, 20). Because ofthe wide distribution of this oncofetal antigen among differentcancer types and its role in in vitro immunodiagnosis (31), CEARAID may prove of significant value in the management ofdiverse cancers showing elevated quantities of this antigen,especially gastrointestinal and mucinous ovarian neoplasms (18,22). However, additional studies are needed to reveal the mostsuitable radiolabel, scanning method and instrumentation, andform of antibody for imaging.

Clinical studies with 131l-labeled anti-CEA IgG have claimed

CANCER RESEARCH VOL. 46 JANUARY 1986

272

Research. on August 30, 2021. © 1986 American Association for Cancercancerres.aacrjournals.org Downloaded from

RADIOANTIBODY WHOLE-BODY AUTORADIOGRAPHY

Table 1Concentration of â„¢5I-anti-CEAIgG and 125/-/gGin hamsters bearing heterotransplanted GW-39 tumors

Quantitation by digitized videodensitometry using 125Ireferencestandards as described in the text. Valuesshown are integrated densitiesexpressed as dpm/mm2for

two subjects per time interval.

TumorBloodLungLiverKidneySpleenMuscle1

day86.5±9.2a' "

79.0 ±9.097.0 ±11.027.3 ±7.549.1 ±4.250.1 ±7.16.1 ±5.7125l-Anti-CEA

IgG3

days118.8

±19.734.4 ±10.831.2 ±8.210.9 ±3.021.6 + 5.014.0 ±2.02.5 ±2.17

days80.5

±14.28.3 ±2.47.5 ±0.73.1 ±0.15.9 ±0.23.9 ±1.20.9 ±0.21

day79.0

±47.376.0 ±12.073.0 ±12.027.6 ±10.755.8 ±5.351.6 ±22.012.0 ±11.2125l-lgG3

days48.0

±9.242.1 ±4.229.0 ±0.02

8.6 ±0.621.0 + 3.014.0 ±4.02.7 ±2.37

days23.2

±6.88.3 ±2.48.0 ±1.43.7 ±3.86.5 ±1.17.3 ±1.21.3 ±1.4

a Mean ±SD.6 Values for tumor radioactivity uptake derived from highest densities (proliferationzones) combined with lowest densities (reducedor non-proliferativezones).

Table 2Videodensitometrically derived tumor ¡non-tumortissue ratios obtained following injection of *xl-anti-CEA IgG

and '25/-/gG

Ratios were obtained from the data presented in Table 1.125l-Anti-CEAIgG1

dayTumor/blood

Tumor/lungTumor/liverTumor/kidneyTumor/spleenTumor/muscleA"0.8

0.62.21.21.2

10.0B1.4

1.14.12.32.2

18.43

daysA2.2

2.46.93.55.3

29.9B4.7

5.214.97.5

11.665.17

daysA6.5

7.217.5

9.213.960.2B12.9

14.234.518.127.4

118.71

dayA0.9

1.02.61.31.46.0B1.1

1.23.11.51.77.2125l-lgG3

daysA1.0

1.44.61.92.8

14.7B1.31.9

6.62.74.0

20.97

daysA1.3

1.43.01.71.58.4B4.3

4.49.65.54.9

27.3a A, ratios derived from lowest densitometric values obtained, corresponding to tumor regions with low to

absent growth and radioactivity accumulation; B, ratios derived from highest densitometric values obtained,corresponding to tumor regions with active proliferation and the most intense radioactivity accumulation.

TablesTumor/non-tumor tissue ratios derived by direct tissue counting

Time post-injection24h72h7

daysNon-tumor

tissueBloodLungs

LiverKidney

SpleenBloodLungs

LiverKidney

SpleenBloodLungsLiverKidneyoplGGnTumor/non-tumor

ratiosGoat

anti-CEA0.7

+0.1"1.5

±0.52.2 +0.31.4

±0.11.7 + 0.33.3 +0.54.8

±0.611.4 +2.27.8

±1.312.5 ±2.14.8 + 1.1

10.5 ±2.021.3±4.512.9

±2.218.5 ±4.2Normal

goatIgG0.4

+0.050.6±0.05

1.7 +0.101.3±0.04

1.7 + 0.210.8 +0.11.5

±0.23.9 ±0.32.8

±0.24.8 ±0.70.8 ±0.12.3 ±0.24.5 ±0.42.8

±0.35.1 ±0.5

a Mean ±SE (n = 6).

tumor regression with therapeutic doses (32), which is alsoindicated by experimental radioimmunotherapy with CEA antibodies in the GW-39 human colonie carcinoma-hamster hostsystem (33). These results suggest an accretion of antibody-mediated radioactivity on or in the CEA-containing tumor cells,

but evidence supporting this has been ambiguous. Whereasprevious autoradiographic studies with anti-CEA polyclonal antibodies did not demonstrate tumor-cell localization (7, 9), a morerecent study with anti-CEA monoclonal antibodies given to mice

bearing human colorectal cancer xenografts indicated that certain monoclonal antibodies resulted in heavily labeled tumor cells(8). In contrast, the current findings clearly support, by micro

scopic autoradiography, the specific accretion of radioactivitygrains in or on the xenografted tumor cells of hamsters receivingaffinity-purified goat anti-CEA IgG, whereas control IgG showed

a reduced and more uniform distribution throughout the tissues.We conclude, therefore, that the inability to demonstrate polyclonal anti-CEA antibody uptake by autoradiographic methodsreported earlier (7, 9) may be related to the use of low-reactiveantibody preparations or of tumors with a low density of availableCEA sites. Although the limitations of autoradiography do notpermit us to conclude whether the selective grain accumulationis on or in the tumor cells, it is interesting to note that previouscell culture studies have shown that CEA antibodies can beretained on the surface of colorectal cancer cells (34-36) and

that the antibodies may be taken into the cells by endocytosis(36).

The WBAR results confirm the tissue counting findings of thecurrent and previous experimental studies (2, 5, 6, 11) thatdemonstrated a specific tumor accretion of radiolabeled anti-

CEA antibodies. In contrast to tissue counting data, where meancpm per tissue weight are expressed, quantitative videodensi-tometric determinations of the tumors revealed a nonuniformdistribution of radioactivity, with outer zones of high activity andcentral zones of low activity. The peripheral zones showed, bymicroscopic autoradiography, intact tumor cells with a concentration of overlying grains, whereas the central portions had areduced and more uniform distribution of grains within areas ofnecrotic cells and stromal connective tissue. If tumor-to-nontar-

get ratios are determined with tumor regions having highest graindensities, then localization indices would far exceed (by almost4-fold) those achieved by using traditional mean tissue counting

CANCER RESEARCH VOL. 46 JANUARY 1986

273

Research. on August 30, 2021. © 1986 American Association for Cancercancerres.aacrjournals.org Downloaded from

RADIOANTIBODY WHOLE-BODY AUTORADIOGRAPHY

data. These results further suggest that tumor size and, in turn,ratios of viable to necrotic cells may influence the accretion ofantitumor antibodies. Alternatively, extensive necrosis may beresponsible for the retention of non-antibody immunoglobulins in

tumors, since clearance mechanisms may be impaired. This viewis consistent with our very early observations in 1974, when wereported the increased localization of normal goat IgG in xeno-grafted human GW-39 tumors as compared to normal hamster

host organs (2). Subsequently, we found that patients with largetumors could be imaged with radioiodinated normal goat IgG(20). These findings caution that imaging relatively large tumorsin patients need not be due to exogenous antibody complexingwith tumor antigen, but instead the nonspecific absorption andretention of macromolecules by neoplasms. Quantitative WBARstudies of tumors of various sizes and necrosis receiving radio-

labeled irrelevant immunoglobulins need to be undertaken tohelp resolve this important issue.

Another factor that may influence the uptake of antibody bythe tumor is the accessibility of the circulating antibody to thetumor cells. Thus, the high concentration of grains on the periphery of the tumor may have been due to a higher concentrationof antigen in these cells, or it may represent a greater accessibilityof the tumor cells to the circulating antibody due to the tumorcells' closer proximity to blood vessels that are more predomi

nantly found at the perimeter of the tumor. Studies that willsimultaneously measure qualitative antigenic expression, tumorviability, and radioantibody uptake are in progress.

These experiments have shown that external tissue countingor imaging of animals receiving radiolabeled antibodies providesonly a gross depiction of relative accretion of radioactivity indifferent regions of the body. The combined use of WBAR andmicroautoradiography further allows clearer interpretations oforgan, tissular, and cellular distribution of radioactivity and howthis may be influenced by a number of tumor and host factors,not least of which are tumor size, tumor morphology, cell viability,antigen density, and microcirculation. These factors must bebetter understood before optimal conditions for improved RAIDof cancer can be defined and achieved.

REFERENCES

1. Colcher, p., Zalutsky, M., Kaplan, W., Kufe, D., Austin, F., and Schlom, J.Radiolocalizationof humanmammarytumors in athymic mice by a monoclonalantibody. Cancer Res., 43: 736-742,1983.

2. Goldenberg,D. M., Preston, D. F., Primus, F. J., and Hansen,H. J. Photoscanlocalization of GW-39 tumors in hamsters using radiolabeled anticarcinoem-bryonic antigen immunoglobulinG. Cancer Res., 34:1-9,1974.

3. Moshakis,V., Mcllhinney,R.A. J., Raghavan,D., and Neville,A. M. Localizationof human tumour xenografts after i.v. administrationof radiolabelledmonoclonal antibodies. Br. J. Cancer,44: 91-99,1981.

4. Moshakis,V., Mcllhinney,R. A. J., Raghavan,D., and Neville,A. M. Monoclonalantibodies to detect human tumours: an experimental approach. J. Clin.Pathol., 34:314-319,1981.

5. Primus, F. J., MacDonald, R., Goldenberg,D. M., and Hansen, H. J. Localization of GW-39 tumors in hamsters by affinity-purified antibody to carcinoem-bryonic antigen. Cancer Res., 37:1544-1547,1977.

6. Primus, F. J., Wang, R. H., Goldenberg,D. M., and Hansen, H. J. Localizationof human GW-39 tumors in hamsters by radiolabeledheterospecific antibodyto carcinoembryonicantigen. Cancer Res., 33: 2977-2982,1973.

7. Lewis, J. C. M., Bagshawe, K. D., and Keep, P. A. The distribution ofparenterallyadministeredantibody to CEA incolorectal xenografts: preliminaryfindings. Oncodevel. Biol. Med., 3:161-168,1982.

8. Lewis, J. C. M., Boxer, G. M., Searle,F., and Bagshawe,K. D.The comparativedistribution of monoclonal antibodies to CEA in colorectal xenografts. TumorBiol., 5: 255-261, 1984.

9. Moshakis, V., Ormerod, M. G., Westwood, J. H., Imrie, S., and Neville,A. M.

The site of binding of anti-CEA antibodies to tumour CEA in vivo: an immu-nocytochemical and autoradiographic approach. Br. J. Cancer, 46: 18-21,1982.

10. Epenetos, A. A., Nimmon, C. C.., Arklie, J., Elliott, A. T., Hawkins, L. A.,Knowles, R. W., Britton, K. E., and Bodmer, W. F. Detectionof humancancerin an animal model using radio-labelledtumour-associated monoclonal antibodies. Br. J. Cancer,46: 1-8,1982.

11. Wahl, R. L., Parker, C. W., and Philpott, G. W. Improved radioimaging andtumor localizationwith monoclonalF(ab')z.J. Nucí.Med., 24: 317-325,1983.

12. Goldenberg, D. M. and DeLand, F. H. History and status of tumor imagingwith radiolabeledantibodies.J. Biol. ResponseModif., 1:121-136,1982.

13. Goldenberg,D. M., DeLand,F., Kim, E., Bennett, S., Primus,F. J., van Nagell,J. R., Jr., Estes, N., DeSimone, P., and Raybum, P. Use of radiolabeledantibodies to carcinoembryonic antigen for the detection and localization ofdiversecancersby external photoscanning.N. Engl.J. Med., 298:1384-1388,1978.

14. Mach, J-P., Buchegger, F., Forni, M., Ritschard, J., Berche, C., Lumbroso, J-D., Schreyer,M., Girardet,C., Accolla,R. S., andCarrel,S. Useof radiolabelledmonoclonal anti-CEA antibodies for the detection of human carcinomas byexternal photoscanning and tomoscintigraphy. Immunol.Today, 1: 239-249,1981.

15. Goldenberg,D. M. Immunodiagnosisand immunodetectionof colorectal cancer. Cancer Bull., 30: 213-218,1978.

16. Goldenberg,D. M. (ed.). Radioimmunodetectionof Cancer workshop. CancerRes., 40: 2957-3087,1980.

17. DeLand, F. H., Kim, E. E., Casper, S., Corgan, R. L, Primus, F. J., Dine, M.E., and Goldenberg, D. M. Radioimmunodetectionof occult recurrent coloniecarcinoma.Am. J. Roentgenol., 738: 145-148,1982.

18. Goldenberg, D. M., Kim, E. E., Bennett, S. J., Nelson, M. 0., and DeLand, F.H. CEA radioimmunodetectionin the evaluationof colorectal cancer and in thedetection of occult neoplasms.Gastroenterology,84: 524-532,1983.

19. Goldenberg,D. M., Kim, E. E., and DeLand,F. H. Humanchorionic gonadotro-pin radioantibodies in the radioimmunodetectionof cancer and for disclosureof occult métastases.Proc. Nati. Acad. Sci. USA, 78: 7754-7758,1981.

20. Goldenberg, D. M., Kim, E. E., DeLand, F. H., Bennett, S., and Primus, F. J.Radioimmunodetectionof cancer with radioactive antibodies to carcinoembryonic antigen. Cancer Res., 40: 3008-3012,1980.

21. Hatch, K. D., Mann, W. J., Boots, L. R., Tauxe, W. N., Shingleton, H. M., andBuchina, E. S., Jr. Localization of choriocarcinoma by 131I-/3HCGantibody.Gynecol.Oncol., 70: 253-261,1980.

22. Van Nagell,J. R., Jr., Kim, E., Casper, S., Primus, F. J., Bennett, S., DeLand,F. H., and Goldenberg,D. M. Radioimmunodetectionof primaryand metastaticovarian cancer using radiolabeled antibodies to carcinoembryonic antigen.Cancer Res., 40: 502-506, 1980.

23. Goldenberg, D. M. and Hansen, H. J. Carcinoembryonic antigen present inhuman colonie neoplasms serially propagated in hamsters. Science (Wash.DC), 775:1117-1118,1972.

24. Goldenberg, D. M., Witte, S., and Elster, K. GW-39: a new human tumorserially transplantable in the golden hamster. Transplantation (Baltimore), 4:760-763,1966.

25. Greenwood, F. G., Hunter, W. M., and Glover, J. S. The preparation of 131I-labeledhumangrowth hormoneof high specific radioactivity. Biochem.J., 89:114-123,1963.

26. Fand, I. and McNally,W. P. The techniqueof whole-body autoradiography. In:J. E. Johnson, Jr. (ed.), Current Trends in MorphologicalTechniques, Vol. 2,pp. 1-28, Boca Raton, FL: CRC Press, Inc., 1981.

27. Ullberg,S. Studies on the distribution and fate of ^S-labeled benzylpenicillininthe body. Acta Radiol. (Suppl.), 778: 1-110, 1954.

28. Hendrickson, A. Light microscopic autoradiography by biological specimens.In: J. E. Johnson (ed.), Current Trends in Morphological Techniques, Vol. 2,pp. 29-52. Boca Raton, FL: CRC Press, Inc., 1981.

29. Rogers, A. W. Techniques of Autoradiography. New York: Elsevier/North-Holland BiomedicaiPress, 1979.

30. Cross, S. A. M., Groves, A. D., and Hesselbo, T. A quantitative method formeasuring radioactivity in tissues sectioned for whole body autoradiography.Int. J. Appi. Radiât.Isotop., 25: 381-386,1974.

31. Goldenberg,D. M., Neville,A. M., Carter, A. C., Go, V. L. W., Holyoke, E. D.,Isselbacher, K. J., Schein, P. S., and Schwartz, M. CEA (carcinoembryonicantigen): its role as a marker in the management of cancer. J. Cancer Res.Clin. Oncol., 707: 239-242,1981.

32. Order, S. E., Klein,J. L., Ettinger, D., Alderson,P., Siegelman,S., and Leichner,P. Use of isotopie immunoglobulin in therapy. Cancer Res., 40: 3001-3007,1980.

33. Goldenberg,D. M., Gaffar, S. A., Bennett, S. J., and Beach,J. L. Experimentalradioimmunotherapyof a xenograftedhumancolonietumor (GW-39)producingcarcinoembryonicantigen. Cancer Res., 41: 4354-4360,1981.

34. Coates, J. E., Koch, M., Beaver, P. F., McPherson,T. A., and Noujaim, A. A.Radioiodinatedantibody to carcinoembryonicantigen: binding to normal andcancerous humancolon in vitro. J. Nati. Cancer Inst, 55: 25-27, 1975.

35. Rosenthal, K. L, Palmer,J. L., Harris, J. A., Rawls, W. E., and Tompkins, W.A. F. Antibody-inducedredistribution of CEA on the cell surface: utilization inseparationof CEA and isoantigenA. J. Immunol., 775: 1049-1053,1975.

36. Rosenthal, K. L., Tompkins, W. A. F., and Rawls, W. E. Factors affecting theexpressionof carcinoembryonicantigenat the surfaceof cultured humancoloncarcinomacells. Cancer Res., 40: 4744-4750,1980.

CANCER RESEARCH VOL. 46 JANUARY 1986

274

Research. on August 30, 2021. © 1986 American Association for Cancercancerres.aacrjournals.org Downloaded from

RADIOANTIBODY WHOLE-BODY AUTORADIOGRAPHY

CANCER RESEARCH VOL. 46 JANUARY 1986

275

Research. on August 30, 2021. © 1986 American Association for Cancercancerres.aacrjournals.org Downloaded from

RADIOANTIBODY WHOLE-BODY AUTORADIOGRAPHY

CANCER RESEARCH VOL. 46 JANUARY 1986

276

Research. on August 30, 2021. © 1986 American Association for Cancercancerres.aacrjournals.org Downloaded from

RADIOANTIBODY WHOLE-BODY AUTORADIOGRAPHY

?s••-.'r ?•&'•.«*,¿fr*^Ä

e*-

^r-

• - '•'*

.% -'¿.'%

Fig.4. Microautoradiographic localization of radiolabeled anti-CEA antibody in formalin-fixed, paraffin-embedded sections of GW-39 tumors counterstained withhematoxylin-eosin:a, enriched uptake of antibody in the cells located toward the periphery (P)of the tumor (magnification,x 120); b, higher magnificationshowing theselective uptake of radioantibody at the tumor cells (7) located in pseudoacinicstructures. Stromal areas (Sf)are relatively free of antibody activity (magnification,x 300).A blood vessel is noted in the upper right-hand comer (BV).

CANCER RESEARCH VOL. 46 JANUARY 1986

277

Research. on August 30, 2021. © 1986 American Association for Cancercancerres.aacrjournals.org Downloaded from

1986;46:271-277. Cancer Res   Irwin Fand, Robert M. Sharkey, William P. McNally, et al.   Antibody Distribution in a Xenografted Human Cancer ModelQuantitative Whole-Body Autoradiography of Radiolabeled

  Updated version

  http://cancerres.aacrjournals.org/content/46/1/271

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

  .pubs@aacr.orgDepartment at

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/46/1/271To request permission to re-use all or part of this article, use this link

Research. on August 30, 2021. © 1986 American Association for Cancercancerres.aacrjournals.org Downloaded from