Monoclonal Antibodies Reactive With Small Cell
`Carcinoma of the Lung 1·z3
`
`Edward D. Ball,4·s.s Robert F. Grazlano,s Ollve S. Pettenglll,7
`George D. Sorenson,7 and Michael W. Fanger4·5
`
`ABSTRACT-Murlne monoclonal antibodies (MoAb) reactive with
`
`lung tumor cell line (isolated from pleural fluid); A549,
`antigens associated wlth small cell carcinoma of the lung (SCCL)
`an adenocarcinoma lung tumor cell line (obtained from
`
`
`were prepared and partially characterized. Four were selected for
`ATCC) (8); Ca Lu-I, a squamous cell lung carcinoma
`
`further study on the basis of their lack of reactivity with normal
`line (ATCC); SK-Lu-I, a lung adenocarcinoma cell line
`(obtained from Dr. J. Fogh, Sloan Keuering Cancer
`
`leukocytes and erythrocytes. These MoAb, designated SCCL-41,
`SCCL-114, SCCL-124, and SCCL-175, are all lgM immunoglobu­
`Institute) (9); BeWo, a choriocarcinoma cell line
`lins. The binding of these MoAb to patient-derived SCCL tumor
`
`(ATCC); DLD-1, a colon carcinoma cell line (provided
`cells, SCCL cell lines, and non-SCCL cell lines was studied by
`by D. Dexter, Providence, R.l.) (JO); and HE-lung,
`
`indirect immunofluorescence and flow cytometry. Considerable
`embryonic lung fibroblasts (obtained from M.A. Bio­
`
`heterogeneity in the expression of these cell surface antigens was
`products, Walkersville, Md.). Ca Lu-I was cultured in
`
`noted among both the patient-derived tumor cells and the SCCL
`McCoy's 5-A medium (GIBCO, Grand Island, N.Y.)
`cell lines. One of the MoAb, SCCL-175, reacted with 7 of 7
`with 10% FBS. A549, SK-LU-I, IMR 32, and HE-lung
`tumor cell samples and 9 of 10 SCCL cell lines.
`patient-derived
`were cultured in Dulbccco's m<Xlified Eagle minimum
`None of the antigens defined by these MoAb were expressed on
`essential medium with 103 FBS (HE-lung; 163 FBS);
`non-SCCL lung tumor cell lines. SCCL-175 reacted with cells
`BeWo, DLD-1, DMS 485, and DMS 351 were all
`from both a choriocarcinoma and a colon carcinoma cell line,
`cultured in RPMl-1640 with 20% FBS.
`whereas the other 3 MoAb were unreactive
`with these and several
`The following leukemia cell lines were also studied;
`other tumor cell lines. These MoAb may be useful in the
`HL-60, a promyelocytic leukemia cell line (obtained
`of SCCL tumors.-JNCI 1984;
`diagnosis and subclassification
`from R. C. Gallo, National Institutes of Health,
`.Bethesda, Md.) (/I); K562, an undifferentiated myeloid
`72:593-598.
`leukemia cell lin<' (/2); Daudi. and Epstein-Barr virus­
`transformcd B-cell line (/J); and CCRF-CEM, a T-cell
`leukemia line (/4). These lines were all cultured in
`RPMI-1640 with 20% FBS.
`Patient cells.-Tumor cells from patients with
`SCCL were obraincd from either surgical biopsy
`specimens (3 parients) or at autopsy performed within 1
`hours of death (3 parients). Cells freshly isolated from
`
`SCCL is a heterogeneous disease in which several
`morphologic subclasses of tumor cells have been identi­
`fied (I, 2). In addition, SCCL tumors often contain foci
`of non-SCCL lung tumor cells (3, 4). To study these
`and other parameters of heterogeneity in SCCL wi�h
`specific probes, as well as to develop improved abilities
`to diagnose and m•at this disease, we have prepared
`MoAb reactive with cells from SCCL tumors. Cells
`from patients with SCCL and cell lines derived from
`patients with SCCL were found LO be reactive with
`these MoAb. In this paper we describe the preparation
`and specificity of these MoAb.
`
`ABBREVIATIONS USED: ATCC=Arnl'riran Typ<• Cullure C-Olle(tion;
`AZ=so<liurn ;11idt·; RSA=bovinr S<'rum albumin; FBS=frtal bovin(•
`S('rum: M oAb=monodonal antibody (antibodies); PBS=phosphatt'·
`bufkrrd salitH': RIA=rndioimmtmoassay;
`SCCL=small cell carrinoma
`of thr lung.
`
`MATERIALS AND METHODS
`
`Cell /ines.-Cell lines studied included DMS 44, 47,
`53, 79, 153, 187, 235, 406, 431, and 483, all of which
`were derived from patients with SCCL and have pheno­
`typic characteristics of SCCL (5-7). These cell lines
`were cultured in either Waymouth's MB 752/I medium
`containing 20% FBS (DMS 44, 53, 153, 187, 235, 406,
`431, and 483) or RPMJ-1640 medium with 203 FBS
`(DMS 79).
`Non-SCCL tumor cell lines studied included IMR 32,
`a neuroblastoma line (obtained from the ATCC, Rock­
`ville, Md.); OMS 351, a malignant melanoma cell line
`(isolated from lymph n<Xlc biopsy specimen); Squ Ca, a
`squamous cell lung carcinoma cell line (provided by K.
`Havemann and C. Gropp, Marburg, Federal Republic
`of Germany); DMS 485, a large cell undifferentiated
`
`1 Rt•c(·ivt'CI junr 16, 1983: accl'pt<'CI Novcmbrr 8, 1983.
`1 Supported in p<trl by Public I kahh Savin· (Pl IS) gram s
`CA-31918, CA-31888. and CA-25845 from thC' ="atiooal Cancrr
`lmti111tt· and by PllS gram Al·l9053 from lhC' National lmtiw1r of
`Allt'l'l':Y and lnft•nious Disrascs. ThC' Cytofluorogr.iph was 1h1·
`11:rm·rous gih of tht· Fannie E. Ripp<'I Foundation and is partially
`suppotrr<l b) the Non is C.ouon CanCt>r Ontn corr gr.int CA-23108.
`'This s111<l)' was prC'senwd in part al 1hr annual mt'<'cing of the·
`Amrrican F1-c:kr:t1ion for Clinirnl Rc·search. Washington, D.C ..
`May 2. 1983. and appr<1rr<l in abstr.tCl form in Clinical RC'sear<"h.
`Vol.
`31, No. 2. 1>.
`402A. 1983.
`Dartmouth Mc-dic-.il
`School. Hano\'l·r.
`• O<>pa1 tlll<'nt of �1r<lici1w,
`N.11. 03756.
`s l)(>parum·nt of Minobiology. Dartmouth Medical School.
`
`• Adduss rtpri111 rt'qul'Sts to Dr. Ball, Orp;mmt'nr of Microbiology,
`
`Dartmouth Mt'dic<tl School.
`of Pathology, Dartmouth M c-dirnl &hoo f.
`1 Ot'lxirlm<·m
`
`593
`
`JNCl. VOL. 72. NO. 3. MARCH 1981
`
`1 of 6
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`BI Exhibit 1006
`
`

`

`594 Ball, Graziano, Pettengill, et al.
`
`RESULTS
`
`terminal pentasaccharide, lacto-N-fucopentaose III, to
`which several reported anti-SCCL MoAb react (17-19),
`we performed inhibition studies. Purified LNF III
`(provided by V. Ginsburg, National Institutes of
`Health, Bet.hesda, Md.) was incubated with each of the
`MoAb and a known positive control MoAb reactive
`with LNF III, PM-81 (20), at a concentration of 5
`mg/ml for 1 hour before addition of these mixtures lo
`SCCL tumor cells from patient B (text-fig. 2). Quanti­
`tative comparisons of the binding of each MoAb either
`in the presence or absence of LNF III was determined
`by indirect immunofluorescence and flow cytometry.
`
`primary or metastatic tumors, which in all cases were
`densely involved with SCCL, were gently Leased apart
`into single-cell suspensions and passed through a
`stainless-steel filter. Cell viability was assessed by stain­
`ing with acridine orange and ethidium bromide. Only
`samples with more than 50% viable cells were included
`in this study.
`Normal cells.- Normal leukocytes were obtained
`from volunteers and separated into granulocyte, mono­
`cyte, and lymphocyte fractions as previously described
`(15). Erythrocytes typed with antisera to Lewis A and B
`blood group antigens were obtained from the Blood
`Bank of Mary Hitchcock Memorial Hospital, Hanover,
`N.H.
`Preparation of hybridomas.-BALB!c mice were
`immunized ip three times over a 3-month period with
`of hybridomas.-The reactivity of the 4
`Specificity
`2XI07 cells, which had been dissociated from the
`MoAb with the immunogen is shown in text-figure 1.
`primary lung tumor of a patient with SCCL. This
`Intense but variable [Juorescence was observed with
`tumor was classified as "intermediate" in the modified
`each MoAb. None of the MoAb reacted at all by
`World Health Organization classification (2). Fusion of
`indirect immunofluorescence wilh lymphocytes, mono­
`spleen cells from an immunized mouse with murine
`cytes, or granulocytes from the patient from whom the
`myeloma cells of the NS-I cell line was performed with
`primary tumor was obtained or from 6 normal donors
`(data not shown), thus indicating that they are probably
`the use of polyethlene glycol as the fusing agent as
`previously described (/6). Hybridomas making MoAb
`not react.ive with histocompatibility or other common
`reactive with the immunogen were selected by solid­
`cell surface antigens. In addition, none of the MoAb
`phase RIA with the use of glutaraldehyde-fixed cells as
`reacted with erythrocytes from 4 normal donors (2
`previously described (15). Of these, 4 were selected for
`Lewis B and I Lewis A-positive, and I Lewis antigen­
`more extensive study on the basis of their relative
`negative). The reactivities of these MoAb with cells
`from the primary lung tumor of a second patient with
`specificity for the immunogen and lack of reactivity
`SCCL are shown in text-figure 2. In addition, liver
`with normal leukocytes. These hybridomas were desig­
`nated SCCL-41 , SCCL-114, SCCL-124, and SCCL-175.
`metastases from the same patient were examined and
`All 4 of the MoAb produced were IgM antibodies.
`found to have a similar antigenic profile, although
`staining was less intense in each case. A summary of
`
`Indirect immunof luorescence and cytoflttorographic
`analysis.-The reactivity of these MoAb with SCCL
`the reactivities of the MoAb with tumor cells obtained
`cells was determined by indirect immunofluorescence
`from a total of 7 sites from 6 different patients is shown
`and flow cytometry. Cells freshly isolated from primary
`in table I.
`or metastatic tumors were prepared as described above.
`with SCCL cell lines.-The expression of
`Reactivity
`Adherent cell lines were dissociated from the culture
`these antigens on several SCCL cell lines developed by
`flask after IO-minute incubation with 0.01% £OTA
`Pettengill et al. (5) was evaluated. Consistent with
`disodium in calcium and magnesium-free Hank's
`other evidence of phenotypic heterogeneity in these
`balanced salt solution followed by washing with
`cell lines, including differential peptide hormone secre­
`RPMl-1640. Cells were incubated for 30 minutes at 4°C
`tion (6, 21, 22) and morphology (5), we have found
`with 0.5 ml hybridoma supernatant, unbound MoAb
`considerable heterogeneity in antigen expression. As
`were removed by washing wilh 7 volumes of PBS (pH
`examples, the reactivities of MoAb SCCL-124 and
`7.4) containing 0.13 BSA and 0.053 AZ followed by the
`SCCL-175 with 4 differem SCCL cell lines are shown
`addition of fluorescein isothiocyanate-labeled F(ab')2
`in text-figures 3 and 4. Marked heterogeneity in the
`goat anti-mouse antibody (Boehringer-Mannheim,
`expression of antigens defined by each MoAb was
`Indianapolis, Ind.) for an additional 30 minutes. After
`noted.
`another wash in PBS-BSA-AZ, the cells were analyzed
`The pattern of reactivity of these MoAb with 10
`for fluorescence on the Cytofluorograph 50H (Onho
`different SCCL cell lines is shown in table 2. Interest­
`Instruments, Westwood, Mass.) with the 2150 computer
`ingly, this pattern of reactivity is similar to that
`system. Simultaneous gating on both viable and single­
`obtained with fresh tumor cells from patients. For
`cell populations was performed, and the data reported
`example, SCCL-175 reacted significantly with all of the
`are those obtained from these populations. Positive
`samples of fresh tumor tissue as well as the cell lines.
`control antibodies included an MoAb to beta-2-micro­
`In contrast, SCCL-4 I reacted with only 4 of the 7 fresh
`globulin, AML-1-201, and a mouse antiserum obtained
`tumor samples and with 3 of the 10 SCCL cell lines.
`by immunization with fresh SCCL cells. The negative
`Reactivity with non-SCCL tumor cell Lines.-To
`control antibody was an IgM MoAb, SCCI, reactive
`further study the specificity of these MoAb, we exam­
`with an irrelevant antigen (sheep erythrocytes).
`ined a number of lung tumor cell lines of non-SCCL
`studies.-To determine if any of
`Hapten inhibition
`histology, other solid tumor cell lines, and leukemia
`these MoAb react with molecules that possess the
`cell lines. None reacted with cells from several non-
`
`JNCI. VOL. 72, NO. 3. MARCH 1981
`
`5
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`BI Exhibit 1006
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`

`to Small Cell Lung Carcinoma 595
`Antibodies
`
`TEXT-FICllRE
`1.-Reaciivity of 4 MoAb
`
`with patiem A-derived SCCL cells.
`Cells from the primary lung tumor
`
`1hat were used as the immunogen for
`these hybridomas were studied by cyto·
`
`
`fluorography. The specific staining or
`tumor cells (black area) for each MoAb
`is shown in individual panels (A.
`SCCL-41: B. SCCL-114; C, SCCL-124;
`D, SCCL-17"1). The percentage or
`posi1ive cells, as defined by fluores·
`cence greaier than on control MoAb·
`treated cells (dotted line), is shown as
`
`
`well as the mean fluorescence intensity
`(MFI). ll1e vertical line al tile right of
`each panel repre$en is highly fluores­
`cent cells.
`
`PATIENT A: PRIMARY LUNG TUMOR
`
`A
`
`c
`
`SCCL 41
`62% +
`MFI 69
`
`SCCL 124
`64% +
`MFI 75
`
`,, ,
`I
`: I
`I I
`I I
`I \
`a:
`w
`' \
`ID
`:Ii
`::::>
`z
`..J
`..J
`w
`(.)
`
`8
`
`SCCL. 114
`6()% +
`MFI 35
`
`/\
`I I
`I I
`I
`I
`
`D
`
`'1
`I I
`I I
`
`I \
`I \
`I \
`\
`
`SCCL 175
`77% +
`MFI 127
`
`FLUORESCENCE INTENSITY
`
`PATIENT B: PRIMARY LUNG TUMOR
`
`A
`
`c
`
`SCCL 41
`43% +
`MFI 21
`
`SCCL 124
`21% +
`MFI 8
`
`EXT·t-IGllRE
`2.-Re�ctivity of 4 MoAb
`
`with patient B-derived SCCL a-lls.
`Cells obqlined frorn th� prim<iry lung
`tumor of a patient with SCCL weri­
`studied by <"ytofluorogrnphy.
`See kgend
`I for details.
`for text-figure
`
`a:
`w
`ID
`:Ii
`::::>
`z
`..J
`..J w
`(.)
`
`8
`
`SCCL 114
`22% +
`MFI 9
`
`D
`I
`I
`I
`I
`I
`I
`I
`I
`I
`I
`
`SCCL 175
`64% +
`MFI 35
`
`FLUORESCENCE INTENSITY
`
`JNCI, VOL. 72. NO. 3, MARCH 1984
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`3 of 6
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`

`

`596 Ball, Graziano, Pettengill,
`et al.
`
`A Bl B2 c D
`SCCL-41 62 43 42 16 15
`SCCL-114 60 22 12 5 22
`SCCL-124 64 21 8 22 23 +
`SCCL-175 77 64 36 36 53 +
`
`+
`+
`+
`+
`
`TABLE 1.-Reactfoity of MoAb with SCCL tumor cells
`
`b
`freshly isolated from patients0•
`
`Percent positive cells
`Patient Patient
`for patient
`E
`F
`
`MoAb
`
`SCCL lung tumor cell lines of epidermoid, adeno­
`carcinoma, and large cell phenotype (table 3). Only
`other tumor cell lines
`SCCL-175 reacted with cells from
`and colon carcinoma, table 3). None
`(choriocarcinoma
`of the MoAb reacted with cells from the leukemia cell
`lines K562, HL-60, Daudi, and CCRF-CEM (table 3).
`
`Finally, none of the MoAb reacted with HE-lung
`fibroblasts.
`Hapten inhibition studies.-The
`
`binding of each
`MoAb to SCCL tumor cells was not inhibited by
`a Reactivity of MoAb with SCCL cells was determined by
`
`of purified LNF III that have been
`concentrations
`
`indirect immunofluorescence and flow cytometry for patients
`completely binding of LNF III·
`shown to inhibit
`
`
`A-D. The numbers reported for these patients' cells are the
`reactive MoAb (17) and to abolish completely the
`
`percentage of cells stained with spedCic MoAb fluorescing
`
`binding of the PM-81 MoAb. The percentages of
`
`greater than cells stained with control lgM MoAb. Cells from
`
`patient E were examined by indirect immunofluorescence of
`
`positive cells and their mean fluorescence intensities
`
`
`tissue sections and patient F. by RIA. In the latter case. positive
`2
`
`were nearly identical to those shown in text-figure
`
`
`(+) indicates a significant reaction over background fluores­
`both in the presence and absence of LNF III.
`
`cence. RIA counts were greater than three times background
`
`and similar to those obtained with an anti-SCCL antiserum.
`bSamples Bl and B2 were :primary lung tumor cells (Bl) and
`DISCUSSION
`
`
`
`liver metastases (B2) from the same patient. Cells obtained from
`patients A. B. and E were obtained at autopsy. Cells from
`Using the approach of immunizing with fresh SCCL
`
`
`patients C. D. and F were obtained by surgical biopsy. Samples,
`
`tumor tissue, we have prepared and partially character­
`A. Bl. C. and F were obtained from the primary lung tumor.
`MoAb that appear to be
`ized 4 hybridoma-derived
`
`samples B2 and E were liver metastases. and sample D was
`
`from a supraclavicular lymph node metastasis.
`specific £or SCCL. Our reason for selecting
`relatively
`
`3.-Reactivity 0£ MoAb
`
`Ti,:XT·t'IGllRE
`SCCL-124 with
`4 DMS SCCL cell lines
`
`by cytofluorography. I A I as determined
`I
`I
`I
`I
`I
`I
`I
`I
`I
`
`SCCL 124
`
`c
`
`OMS 44
`93� +
`
`DMS47
`88% +
`
`cc w
`ID
`:I!:
`:::>
`z
`-'
`_, w
`l)
`
`B
`
`I
`I
`I
`I
`I
`
`0
`
`OMS 53
`39'% +
`
`OMS 406
`13% +
`
`JNCI, VOL 72. NO.
`3, MARCH 1984
`
`FLUORESCENCEINTENSnY
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`4 of 6
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`

`A
`
`SCCL 175
`
`c
`
`to Small Cell Lung Carcinoma 597
`Antibodies
`
`TlXT-t-lCl'RE 4.-Reactivity
`of MoAb
`SCCL· 175 with 4 DMS SCCL cell lines
`
`as studied by cytofluorogmphy.
`
`OMS 47
`17% +
`
`OMS 79
`38% +
`
`a:
`w
`m
`:I!
`:::>
`z
`...J ...J
`w
`t>
`
`OMS 153
`54'% +
`
`I
`I D
`I
`I
`I
`I
`I
`I
`I
`I
`I
`I
`I
`I
`I
`
`OMS 406
`89% +
`
`FLUORESCENCE INTENSITY
`
`This report demonstrates another aspect of the
`
`burg V: Personal communication) and with human
`
`fresh tumor tissue rather than an SCCL cell line as an
`cells known to express molecules with the
`
`immunogen was because of the possibility that a cell
`granulocytes,
`lacto-N .f ucopemaose III to
`terminal pemasaccharide
`line(s) might not mirror the antigen profile of an in
`which many other anti-tumor cell and SCCL MoAb are
`
`vivo tumor. Our approach was successful in that we
`
`The inability of purified LNF III to
`directed (17-19).
`
`were able to produce MoAb reactive with the cells used
`block the binding of each MoAb supports this con­
`
`as immunogen as well as other patient-derived SCCL
`clusion.
`tumor cells and SCCL cell lines. We also showed that
`
`
`
`the MoAb are not reactive with common or histo·
`
`com pa ti bi lity antigens on leukocytes from either the
`patient from whom the tumor immunogen was derived
`of M oAb with ?Wn-SCCL tumor cell lines•·h
`TABLE 3.-Reactivity
`
`or from normal donors. Moreoever, these MoAb do not
`Cell lines
`MoAb
`
`
`react with glycolipids and, in particular, with the
`
`glycolipid molecules with which many reported anti­
`SCCL SCCL SCCL SCCL
`Tissue origin Designation
`tumor cell, including anti-SCCL, MoAb react (17-19).
`41 114 124 175
`
`
`This conclusion is based on the lack of reactivity of our
`Lung. squamous Squ Ca
`
`MoAb with lipid extracts of SCCL tumor cells (Gins-
`Lung, squamous Ca Lu-I
`Lung, large cell DMS 485
`Lung. adenocarcinoma A549
`Lung. adenocarcinoma SK-LU-1
`Neuroblastoma lMR 32
`OMS 351
`Melanoma
`Choriocarcinoma
`BeWo
`Colon carcinoma DLD-1
`
`Promyelocytic leu- HL-00
`kemia
`K562
`Myeloid leukemia
`B-cell leukemia
`Dau di
`T-cell leukemia
`CCRF-CEM
`
`of MoAb u>ith cells of the
`TABLE 2.-Reactil'ity
`DMS SCCL cell lines•
`
`MoAb
`
`DMS SCCL cell linesb
`44 47 53 79 153 187 235 406 431 483
`
`++
`+
`
`+
`+
`SCCL 41
`+ +
`+
`SCCL 114
`SCCL 124 +++ +++ +
`+ +
`+
`+ + ++ + + +++ + +
`SCCL 175 +
`
`+
`
`0Reactivity of MoAb with SCCL cell lines was determined by
`
`of MoAb with cell lines was determined by flow
`flow cytometry.
`•Reactivity
`"Reactions were scored as follows:-. <20% of cells positive;+.
`cytometry.
`b See footnote b. table 2.
`
`20-40% positive: ++, 4()--60% positive:+++. >60% of cells positive.
`
`
`
`JNCI. VOL. 72. NO. 3. MARCH 1984
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`5 of 6
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`

`598 Ball, Graziano, Pettengill, et al.
`
`hecerogeneicy within SCCL tumors. Of considerable
`interest is che similar pattern of reactivity of cells
`freshly isolated from patients and those obtained from
`cell Jines. That each of the 4 MoAb reacted wich a
`similar percentage of both the cell lines and the freshly
`isolated tumor cell samples indicates that che cultured
`cell lines maintain an antigenic profile similar to cells
`in vivo. We are presently studying tumor cells immedi­
`ately after their removal to directly follow changes in
`antigen expression with time.
`An attempt was made to relate the expression of the
`antigens defined by these MoAb to ocher heterogeneous
`phenotypic characteristics of the SCCL cell lines. These
`included morphology (5), ultrastructure (5). hormone
`secretion (6, 21, 22), tissue site of origin
`(5, 7),
`treatment status of the patient (5), growth rate (5, 7),
`and karyotype (Wurster-Hill D.: Personal communica­
`tion.) No meaningful correlations could be made
`between qualitative or quantitative antigen expression
`and any" of these features of the cell lines.
`As noted by ourselves and other investigators, most
`reported tumor cell-reactive MoAb are also reactive
`with other selected tumor types (19, 23, 24) and with
`normal cells of the same or dirferent lineage (15, 16, 19,
`23, 24). SCCL-175 reacted with cells from a colon
`carcinoma line and a choriocarcinoma line. However,
`none of the MoAb reported here reacted with any of 1he
`non-SCCL lung tumor cell lines studied. Although it is
`premature to conclude that any of these MoAb are
`relatively or absolutely specific for SCCL, further study
`of specificity will reveal whether they will be useful in
`distinguishing SCCL from non-SCCL lung tumors. It
`seems likely that combinations of MoAb reported to be
`reactive with non-SCCL lung tumors but not SCCL
`(25, 26) and MoAb specific for SCCL such as those
`reported here might make a useful panel of reagents for
`1he immunodiagnosis of lung tumors.
`In contrast to the leukemias, the study of solid
`tumors is hampered by the difficulties of both obtain­
`ing and utilizing freshly isolated cells from patients.
`Therefore, the use of well-characterized cell lines is of
`great importance to the continued study of cancers such
`as SCCL. Our finding Lhat SCCL cell lines express cell
`surface antigens characteristic of freshly isolated LUmor
`tissue from patients with SCCL further validates 1he
`use of these lines as models of SCCL cell biology.
`Moreover, the potential utility of MoAb in the diag­
`nosis and treatment of this disease may be studied
`utilizing cell lines that mimic the in vivo antigenic
`properties of SCCL tumor cells.
`
`REFERENCES
`
`(/) C\RTF.R D. f.c:ca.f$l"ON JC. Tumors of 1hc lower respiratory
`tract. lo: Atl;is of 1umor pathology, second srrics. fascick 17.
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`6 of 6
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`BI Exhibit 1006
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