- Email: [email protected]
J-chain expression in human cells producing IgG subclasses
CELLULAR
IMMUNOLOGY
J-Chain
91, 5 1S-5 19 (I 985)
Expression
in Human Cells Producing P. L. HABER
Department
IgG Subclasses’
AND J. MESTECKY’
of Microbiology and Division of Clinical Immunology and Rheumatology, Medicine, University of Alabama at Birmingham, Birmingham, Alabama Received
October
29, 1984; accepted
December
Department 35294
of
13, 1984
Previous studies have demonstrated that J chain is expressed not only in cells that produce polymeric immunoglobulins, but also in those engaged in synthesis of monomers including IgG and IgD. The presence of J chain in these cells suggested that its role may not be restricted to the formation of polymers. For the present study, fluorochrome-labeled polyclonal anti-Jchain and monoclonal antibodies to IgG subclasses were used to determine the distribution of J chain in IgG plasma cells from normal human tissues and from pokeweed mitogen (PWM)stimulated human peripheral blood lymphocytes. The results indicate that J chain is not equally distributed among cells producing different IgG subclasses.The percentages of PWM-stimulated cells containing J chain were: 22 + 5 (SE) for IgGl, 49 + 6 for IgG2, 17 + 7 for IgG3, and 64 f 11 for IgG4. Examination of sections of various human lymphoid tissues revealed that the frequency of IgG cells that coexpressed J chain was lower than that observed in the PWM system and displayed variable distribution among IgG subclasses. The frequency of J-chain expression in IgG-producing cells may be related to the degree of cellular maturation and may differ according to the origin of cells. 0 1985 Academic press, IK. INTRODUCTION
In serum and external secretions, J chain is disulfide linked to the polymeric IgA and IgM molecules (for review see (1, 2)). Examination of immunoglobulinproducing cells by immunohistochemical techniques has revealed that this polypeptide is found not only in IgA- or IgM-containing cells, but also in mouse or human cells that express IgG, IgD, H or L chains, or no immunoglobulin chains at all (3-15). Although most of these studies were performed on lymphoblastoid cell lines or on cells from patients with a variety of lymphoproliterative disorders, J chain has also been detected in IgD- and IgG-containing immunocytes from normal or inflammed human tissues (16-l 8) and in pokeweed mitogen (PWM)-stimulated lymphocytes from peripheral blood of normal donors (19-2 1). However, not all IgG-producing cell expressed J chain. To determine whether the expression of J chain in IgG immunocytes is restricted to certain subclasses, monoclonal antibodies to the subclasses of human IgG were used to examine the distribution of J chain in PWMstimulated peripheral blood mononuclear cells (PBMNC) and in spleen, tonsil, lymph node, and intestinal tissues. ’ Supported by U.S. Public Health Source grants AI-10854 and AI-18745. 2 To whom correspondence should be sent at: Department of Microbiology, Medical Center, University of Alabama at Birmingham, Birmingham, Ala. 35294. 515 QOO8-8749185
$3.00
Copyigbt 8 1985 by Academic Press, Inc. All rights of reproduction in any form reserved
516
SHORT COMMUNICATIONS
MATERIALS
AND
METHODS
Cells. PBMNC were isolated from the venous blood of six normal adults by centrifugation on a Ficoll-Hypaque gradient (22). Cells were cultured at a concentration of 1 X lo6 PBMNC/ml in 5- to IO-ml volumes of RPM1 1640 (GIBCO, Grand Island, N.Y.) supplemented with 15% heat-inactivated fetal bovine serum (Reheis Chemical Co., Kankakee, Ill.), penicillin (0.1 mg/ml), streptomycin (100 units/ml), 2.0 mM L-glutamine, and 10 &ml of PWM (GIBCO). Cell cultures were maintained for 7-8 days at 37°C in a moist atmosphere of 95% air and 5% COZ. Cytocentrifuge slides of cell suspensions prepared from human PBMNC and tonsils, lymph nodes, and spleen, obtained from surgery (within 2 hr) and autopsy (within 5 hr postmortem), were fixed in acid alcohol (23) and extensively washed in phosphate (0.01 M)-buffer saline (PBS), pH 7.4, before staining. Intestinal tissues obtained from the same source were frozen in liquid nitrogen, sectioned on a cryotome and fixed in acid-alcohol before staining. Immunochemical reagents. Fluorescein-isothiocyanate (FITC)-labeled rabbit antihuman IgG was purchased commerically (Calbiochem-Behring, San Diego, Calif.) or prepared in this laboratory (24). Anti-human J chain was raised in rabbits immunized with J chain isolated by gel-filtration and ion-exchange chromatography from Ssulfonated polymeric myeloma IgAZ-X protein (25). The monospecificity of this reagent was tested by immunoelectrophoresis, radioimmunoassay (25, 26), electroblotting (14), immunofluorescence and immunoelectron microscopy (8, 15), and immunoprecipitation of biosynthetically-labeled (3SS-methionine) J chain subsequently analyzed by electrophoresis and peptide mapping (13). In all of these tests this reagent reacted with human J chain, but not with any other immunoglobulin component chains. Tetramethyl rhodamine isothiocyanate (TRITC)-labeled anti-J chain was used for the detection of intracellular J chain by immunofluorescence technique (8). Mouse monoclonal antibodies used in these studies to detect cells containing the different human IgG subclasses include: BAM 15 (anti-IgG 1, Seward Laboratories, London); BAM 14 (anti-IgG2, Seward Laboratories); C3-8-34 (anti-IgG3, BectonDickenson, Irvine, Calif,); and 9441~SA (anti-IgG4, BRL, Gaithersburg, Md.). Evidence for the subclass specificity of these antibodies has been reported (27). Affinity-purified FITC-labeled goat anti-mouse immunoglobulin was purchased from Southern Biotechnology Associates (Birmingham, Ala.). Zmmunofluorescence. Appropriately diluted TRITC-labeled anti-J chain was incubated with fixed cells for 45 min and subsequently washed in PBS. Unlabeled monoclonal antibodies to IgG subclasses were then applied for another 45 min followed by FITC-labeled goat anti-mouse Ig (0.1 mg/ml). Dilutions of the monoclonal antibodies and the FITC-labeled goat anti-mouse optimal for immunofluorescence were determined on cytocentrifuge slides of human spleen cells. Stained slides were examined in a fluorescence microscope (Orthomat, Leitz, Wetzlar, W. Germany) with epillumination and narrow band excitation and barrier filters (S525 for FITC and 6 10 for TRITC) with HBO 100 W super pressure mercury lamp as a light source. RESULTS
AND
DISCUSSION
In agreement with previous experiments (19) a large proportion of IgG-containing cells generated by PWM stimulation of PBMNC also expressed J chain. When
SHORT COMMUNICATIONS
517
stained with IgG subclass-specific reagents, J chain could be detected in cells producing any of the human IgG subclasses although the proportion of J-chaincontaining cells was not evenly distributed. Thus, for the cultures from the six individuals studied, J chain was found in 22 + 5% (mean -t 1 SE) of IgGl-, 49 + 6% of IgG2-, 17 If: 7% of IgG3-, and 64 + 11% of IgGCcontaining cells (Table 1). Furthermore, repetitive examinations of PWM-stimulated PBMNC from the same individuals yielded similar results. Compared with PWM-stimulated PBMNC, fewer IgG-containing cells in lymph nodes, tonsils, and spleen expressed J chain. Also, the relative frequency of J-chain expression among the cells containing various IgG subclasses differed from that observed with PWM cultures. In contrast to the high frequency of IgG4 cells that contain J chain in PWM cultures, almost no splenic IgG4 plasma cells expressed J chain, and the frequencies of IgG4 plasma cells expressing J chain in the tonsils and lymph nodes were not higher than that of the other subclasses. The samples of intestine examined contained very few IgG plasma cells but, similar to the PWM cultures, a large proportion of IgG plasma cells contained J chain. Unlike the PWM cultures, however, many IgG3 plasma cells contained J chain while very few of the IgG2 plasma cells did. The higher proportion of J-chain-containing IgG cells among PWM-stimulated PBMNC than among such cells from normal tissues is in agreement with previous studies which suggest that J-chain expression may correlate with the state of cell differentiation: immature IgG blast cells from tonsillar inflammatory lesions or malignant cells from IgG myeloma (5, 6, 8, 10, 11) and lymphoblastoid cell lines (13-l 5) preferentially express J chain. In the PWM-stimulated cultures it is unlikely that such cells are in the process of switching from IgM to IgG production because double staining with anti-IgG and -1gM reagents does not yield cells expressing both classes ( 19). J chain was not, however, evenly distributed among the four IgG subclasses and the relative proportion of J-chain expression among the IgG subclasses varied according to the type of tissue examined. This could possibly result from differences in stimulation of the IgG subclasses. It is intriguing, though, that the increasing order of J-chain expression in PWM cultures was IgG3 < IgGl < IgG2 < IgG4. Although there is no apparent explanation for the correlation, this is also the order of human germ-line gamma chain constant region genes (28) with IgG4 being the most distal from the V.D.J. region. Similar results were obtained with IgAl- and IgA2-containing cells that express J chain in PWM-stimulated PBMNC (unpublished results) or cells from different human tissues (28): a significantly higher number of IgAZpositive cells expressed J chain than IgA 1-positive cells. The intracellular function of J chain remains unclear. In cells that produce polymeric immunoglobulin of IgM and IgA classes, J chain appears to be involved in the process of the assembly of polymeric molecules (for review see (1, 2)). However, in IgG cells, J chain is not secreted and is intracellularly degraded (29). It may be permissible to speculate that the synthesis of J chain, which in malignant cells and lymphoblastoid cell lines precedes the synthesis of immunoglobulin heavy and light chains (13, 15), influences, by an unknown mechanism, expression of IgG subclasses. Interestingly, more J-chain-positive cells are found among IgG and IgA cells that express products of the constant region gene more distal to the V.D.J. region.
’ Not detectable.
12128 2110 l/30
Intestine 1 2 3
121176 4120
2/l 17 3136
node
ISE
61101 7/101 41142 4187
17166 401125 95 1270
(1)
(0)
o/4
(3)
(9) (23)
(
7&l
(8)
(3) (7)
(10)
(67) (53) (47) (53) 49 f 6
(21)
(50)
@)
Cells
(19) (0)
in Plasma
6132 o/20
5158 4117
l/149 l/13
4140 2166 l/14 4153
18/36 18/87 39/58 40176 46197 1181222
J/&G2
Expression
1
(20)
(43)
(2) (‘3)
(20)
(7)
(7) (3) (5) 5+-l
(6)
(32) (35) 22 _t 5
(21) (28)
(12)
21146 21/100
@)
of J-Chain
211175
J/IgG 1
Lymph 1 2
2
Tonsil 1
Mean+-
2 3 4
Spleen 1
PWM-stimulated PBMNC 1 2 3 4 5 6 Mean + 1 SE
Tissue
Frequency
TABLE Producing
10113
317
14126
3142 2113
1 l/142 4150
4198 o/9 1 7178 l/38
12/130 2172 3195 16196 22185 80/178
J&G3
IgG Sub&sses
(54) (43) (77)
(7) (15)
(8) (8)
(4) (0) (9) (3) 4k2
(45) 17 f 7
(26)
(9) (3) (3) (17)
6)
218
15117 217
3145 (ND)
116
2165
l/81 O/18 l/136 (ND) ’
27155 3119 16123 11/14 24130 100/l 10
J&G4
(29) (25)
(88)
(7)
(16)
(70) (79) (80) (91) 64+ 11
(16)
(49)
(96)
ul
g
&
519
SHORT COMMUNICATIONS
ACKNOWLEDGMENTS We thank Mrs. Annette Pitts, Ms. Shirley Prince and Ms. Rose Kulhavy for excellent technical assistance.
REFERENCES ‘1. 2. 3. 4. 5. 6. 7.
8. 9. 10.
Inman, I. P., and Mestecky, J., Contemp. Top. Mol. Immunol. 3, 111, 1974. Koshland, M. E., Adv. Immunol. 20, 41, 1975. Radl, J., Schuitt, H. R. E., Mestecky, J., and Hijmans, W., Adv. Exp. Med. Biol. 45, 57, 1974. Kaji, H. and Parkhouse, R. M. E., Nature (London) 249,45, 1974. Brandtzaeg, P., and Berdal, P., Stand. J. Immunol. 4, 403, 1975. Sarasombath, S., Mestecky, J., and Skvaril, F., Clin. Exp. Immunol. 29, 67, 1977. Isaacson, P., J. Clin. Pathol. 32, 802, 1979. Mestecky, J., Preud’homme, J.-L., Crago, S. S., Mihaesco, E., Prchal, J. T., and Okos, A. J., Clin. Exp. Immunol. 39, 371, 1980. Yasuda, N., Kanoh, T., and Uchino, H., Clin. Exp. Immunol. 40, 573, 1980. Bast, E. J. E. G., vanCamp, B., Boom, S. E., Jaspers, F. C. A., and Ballieux, R., Clin. Exp. Immunol. 44, 375,
1981.
11. Laurent, Cl., Delsol, Cl., Reyes, F., AbbaI, M., and Mihaesco, E., Clin. Exp. Immunol. 44, 620, 1981. 12. Mason, D. Y., and Stein, H., Clin. Exp. Immunol. 46, 305, 198 1. 13. McCune, J. M., Fu, S. M., and Kunkel, H. G., J. Exp. Med. 154, 138, 1981. 14. Kutteh, W. H., Moldoveanu, Z., Prince, S. J., Kulhavy, R., Alonso, F., and Mestecky, J., Mol. Immunol. 20, 967, 1983. 15. Hajdu, I., Moldoveanu, Z., Cooper, M. D., and Mestecky, J., J. Exp. Med. 158, 1993, 1983. 16. Brandtzaeg, P., Nature (London) 252, 418, 1974. 17. Brandtzaeg, P., Gjeruldsen, S. T., Korsrud, F., Baklien, K., Berdal, P., and Ek, J., J. Immunol. 122, 503, 1979.
18. Korsrud, F. R., and Brandtzaeg, P., Stand. J. 19. Mestecky, J., Winchester, R. J., Hoffman, T., 20. Yasuda, N., Kanoh, T., and Uchino, H., Clin. 21. Reitamo, S., K;iyhkG, K., Konttinen, Y. T.,
Immunol.
13, 281,
1981.
and Kunkel, H. G., J. Exp. Med. Exp. Immunol.
and Bergroth,
46, 142, 198 I. V. J. Histochem.
145, 760, 1977. Cytochem.
31, 843,
1983. 22. Boyurn, A., Stand. J. Clin. Lab. Invest. 21 (Suppl. 97), 77, 1968. 23. Hijmans, W., Schuitt, H. R. E., and Klein, F., Clin. Exp. Immunol. 4, 457, 1969. 24. Crago, S. S., and Mestecky, J., J. Immunol. 122, 906, 1979. 25. Mestecky, J., Zikan, J., Butler, W. T., and Kulhavy, R., Immunochemistry 9, 883, 1972. 26. Kutteh, W. H., Prince, S. J., Phillips, J. O., Spenney, J. G., and Mestecky, J., Gastroenterology
81,
184, 1982. 27. Mayumi, M., Kuritani, T., Kubagawa, H., and Cooper, M., J. Immunol. 130, 671, 1983. 28. Flanagan, J. G., and Rabbitts, T. H., Nature (London) 300, 709, 1982. 29. Crago, S. S., Kutteh, W. H., Moro, I., Allansmith, M. R., Radl, J., Haaijman, J. J., and Mestecky, J., J. Immunol. 132, 16, 1984. 30. Mosmann, T. R., Gravel, Y., Williamson, A. R., and Baumal, R., Eur. J. Immunol. 8, 94, 1978.
IMMUNOLOGY
J-Chain
91, 5 1S-5 19 (I 985)
Expression
in Human Cells Producing P. L. HABER
Department
IgG Subclasses’
AND J. MESTECKY’
of Microbiology and Division of Clinical Immunology and Rheumatology, Medicine, University of Alabama at Birmingham, Birmingham, Alabama Received
October
29, 1984; accepted
December
Department 35294
of
13, 1984
Previous studies have demonstrated that J chain is expressed not only in cells that produce polymeric immunoglobulins, but also in those engaged in synthesis of monomers including IgG and IgD. The presence of J chain in these cells suggested that its role may not be restricted to the formation of polymers. For the present study, fluorochrome-labeled polyclonal anti-Jchain and monoclonal antibodies to IgG subclasses were used to determine the distribution of J chain in IgG plasma cells from normal human tissues and from pokeweed mitogen (PWM)stimulated human peripheral blood lymphocytes. The results indicate that J chain is not equally distributed among cells producing different IgG subclasses.The percentages of PWM-stimulated cells containing J chain were: 22 + 5 (SE) for IgGl, 49 + 6 for IgG2, 17 + 7 for IgG3, and 64 f 11 for IgG4. Examination of sections of various human lymphoid tissues revealed that the frequency of IgG cells that coexpressed J chain was lower than that observed in the PWM system and displayed variable distribution among IgG subclasses. The frequency of J-chain expression in IgG-producing cells may be related to the degree of cellular maturation and may differ according to the origin of cells. 0 1985 Academic press, IK. INTRODUCTION
In serum and external secretions, J chain is disulfide linked to the polymeric IgA and IgM molecules (for review see (1, 2)). Examination of immunoglobulinproducing cells by immunohistochemical techniques has revealed that this polypeptide is found not only in IgA- or IgM-containing cells, but also in mouse or human cells that express IgG, IgD, H or L chains, or no immunoglobulin chains at all (3-15). Although most of these studies were performed on lymphoblastoid cell lines or on cells from patients with a variety of lymphoproliterative disorders, J chain has also been detected in IgD- and IgG-containing immunocytes from normal or inflammed human tissues (16-l 8) and in pokeweed mitogen (PWM)-stimulated lymphocytes from peripheral blood of normal donors (19-2 1). However, not all IgG-producing cell expressed J chain. To determine whether the expression of J chain in IgG immunocytes is restricted to certain subclasses, monoclonal antibodies to the subclasses of human IgG were used to examine the distribution of J chain in PWMstimulated peripheral blood mononuclear cells (PBMNC) and in spleen, tonsil, lymph node, and intestinal tissues. ’ Supported by U.S. Public Health Source grants AI-10854 and AI-18745. 2 To whom correspondence should be sent at: Department of Microbiology, Medical Center, University of Alabama at Birmingham, Birmingham, Ala. 35294. 515 QOO8-8749185
$3.00
Copyigbt 8 1985 by Academic Press, Inc. All rights of reproduction in any form reserved
516
SHORT COMMUNICATIONS
MATERIALS
AND
METHODS
Cells. PBMNC were isolated from the venous blood of six normal adults by centrifugation on a Ficoll-Hypaque gradient (22). Cells were cultured at a concentration of 1 X lo6 PBMNC/ml in 5- to IO-ml volumes of RPM1 1640 (GIBCO, Grand Island, N.Y.) supplemented with 15% heat-inactivated fetal bovine serum (Reheis Chemical Co., Kankakee, Ill.), penicillin (0.1 mg/ml), streptomycin (100 units/ml), 2.0 mM L-glutamine, and 10 &ml of PWM (GIBCO). Cell cultures were maintained for 7-8 days at 37°C in a moist atmosphere of 95% air and 5% COZ. Cytocentrifuge slides of cell suspensions prepared from human PBMNC and tonsils, lymph nodes, and spleen, obtained from surgery (within 2 hr) and autopsy (within 5 hr postmortem), were fixed in acid alcohol (23) and extensively washed in phosphate (0.01 M)-buffer saline (PBS), pH 7.4, before staining. Intestinal tissues obtained from the same source were frozen in liquid nitrogen, sectioned on a cryotome and fixed in acid-alcohol before staining. Immunochemical reagents. Fluorescein-isothiocyanate (FITC)-labeled rabbit antihuman IgG was purchased commerically (Calbiochem-Behring, San Diego, Calif.) or prepared in this laboratory (24). Anti-human J chain was raised in rabbits immunized with J chain isolated by gel-filtration and ion-exchange chromatography from Ssulfonated polymeric myeloma IgAZ-X protein (25). The monospecificity of this reagent was tested by immunoelectrophoresis, radioimmunoassay (25, 26), electroblotting (14), immunofluorescence and immunoelectron microscopy (8, 15), and immunoprecipitation of biosynthetically-labeled (3SS-methionine) J chain subsequently analyzed by electrophoresis and peptide mapping (13). In all of these tests this reagent reacted with human J chain, but not with any other immunoglobulin component chains. Tetramethyl rhodamine isothiocyanate (TRITC)-labeled anti-J chain was used for the detection of intracellular J chain by immunofluorescence technique (8). Mouse monoclonal antibodies used in these studies to detect cells containing the different human IgG subclasses include: BAM 15 (anti-IgG 1, Seward Laboratories, London); BAM 14 (anti-IgG2, Seward Laboratories); C3-8-34 (anti-IgG3, BectonDickenson, Irvine, Calif,); and 9441~SA (anti-IgG4, BRL, Gaithersburg, Md.). Evidence for the subclass specificity of these antibodies has been reported (27). Affinity-purified FITC-labeled goat anti-mouse immunoglobulin was purchased from Southern Biotechnology Associates (Birmingham, Ala.). Zmmunofluorescence. Appropriately diluted TRITC-labeled anti-J chain was incubated with fixed cells for 45 min and subsequently washed in PBS. Unlabeled monoclonal antibodies to IgG subclasses were then applied for another 45 min followed by FITC-labeled goat anti-mouse Ig (0.1 mg/ml). Dilutions of the monoclonal antibodies and the FITC-labeled goat anti-mouse optimal for immunofluorescence were determined on cytocentrifuge slides of human spleen cells. Stained slides were examined in a fluorescence microscope (Orthomat, Leitz, Wetzlar, W. Germany) with epillumination and narrow band excitation and barrier filters (S525 for FITC and 6 10 for TRITC) with HBO 100 W super pressure mercury lamp as a light source. RESULTS
AND
DISCUSSION
In agreement with previous experiments (19) a large proportion of IgG-containing cells generated by PWM stimulation of PBMNC also expressed J chain. When
SHORT COMMUNICATIONS
517
stained with IgG subclass-specific reagents, J chain could be detected in cells producing any of the human IgG subclasses although the proportion of J-chaincontaining cells was not evenly distributed. Thus, for the cultures from the six individuals studied, J chain was found in 22 + 5% (mean -t 1 SE) of IgGl-, 49 + 6% of IgG2-, 17 If: 7% of IgG3-, and 64 + 11% of IgGCcontaining cells (Table 1). Furthermore, repetitive examinations of PWM-stimulated PBMNC from the same individuals yielded similar results. Compared with PWM-stimulated PBMNC, fewer IgG-containing cells in lymph nodes, tonsils, and spleen expressed J chain. Also, the relative frequency of J-chain expression among the cells containing various IgG subclasses differed from that observed with PWM cultures. In contrast to the high frequency of IgG4 cells that contain J chain in PWM cultures, almost no splenic IgG4 plasma cells expressed J chain, and the frequencies of IgG4 plasma cells expressing J chain in the tonsils and lymph nodes were not higher than that of the other subclasses. The samples of intestine examined contained very few IgG plasma cells but, similar to the PWM cultures, a large proportion of IgG plasma cells contained J chain. Unlike the PWM cultures, however, many IgG3 plasma cells contained J chain while very few of the IgG2 plasma cells did. The higher proportion of J-chain-containing IgG cells among PWM-stimulated PBMNC than among such cells from normal tissues is in agreement with previous studies which suggest that J-chain expression may correlate with the state of cell differentiation: immature IgG blast cells from tonsillar inflammatory lesions or malignant cells from IgG myeloma (5, 6, 8, 10, 11) and lymphoblastoid cell lines (13-l 5) preferentially express J chain. In the PWM-stimulated cultures it is unlikely that such cells are in the process of switching from IgM to IgG production because double staining with anti-IgG and -1gM reagents does not yield cells expressing both classes ( 19). J chain was not, however, evenly distributed among the four IgG subclasses and the relative proportion of J-chain expression among the IgG subclasses varied according to the type of tissue examined. This could possibly result from differences in stimulation of the IgG subclasses. It is intriguing, though, that the increasing order of J-chain expression in PWM cultures was IgG3 < IgGl < IgG2 < IgG4. Although there is no apparent explanation for the correlation, this is also the order of human germ-line gamma chain constant region genes (28) with IgG4 being the most distal from the V.D.J. region. Similar results were obtained with IgAl- and IgA2-containing cells that express J chain in PWM-stimulated PBMNC (unpublished results) or cells from different human tissues (28): a significantly higher number of IgAZpositive cells expressed J chain than IgA 1-positive cells. The intracellular function of J chain remains unclear. In cells that produce polymeric immunoglobulin of IgM and IgA classes, J chain appears to be involved in the process of the assembly of polymeric molecules (for review see (1, 2)). However, in IgG cells, J chain is not secreted and is intracellularly degraded (29). It may be permissible to speculate that the synthesis of J chain, which in malignant cells and lymphoblastoid cell lines precedes the synthesis of immunoglobulin heavy and light chains (13, 15), influences, by an unknown mechanism, expression of IgG subclasses. Interestingly, more J-chain-positive cells are found among IgG and IgA cells that express products of the constant region gene more distal to the V.D.J. region.
’ Not detectable.
12128 2110 l/30
Intestine 1 2 3
121176 4120
2/l 17 3136
node
ISE
61101 7/101 41142 4187
17166 401125 95 1270
(1)
(0)
o/4
(3)
(9) (23)
(
7&l
(8)
(3) (7)
(10)
(67) (53) (47) (53) 49 f 6
(21)
(50)
@)
Cells
(19) (0)
in Plasma
6132 o/20
5158 4117
l/149 l/13
4140 2166 l/14 4153
18/36 18/87 39/58 40176 46197 1181222
J/&G2
Expression
1
(20)
(43)
(2) (‘3)
(20)
(7)
(7) (3) (5) 5+-l
(6)
(32) (35) 22 _t 5
(21) (28)
(12)
21146 21/100
@)
of J-Chain
211175
J/IgG 1
Lymph 1 2
2
Tonsil 1
Mean+-
2 3 4
Spleen 1
PWM-stimulated PBMNC 1 2 3 4 5 6 Mean + 1 SE
Tissue
Frequency
TABLE Producing
10113
317
14126
3142 2113
1 l/142 4150
4198 o/9 1 7178 l/38
12/130 2172 3195 16196 22185 80/178
J&G3
IgG Sub&sses
(54) (43) (77)
(7) (15)
(8) (8)
(4) (0) (9) (3) 4k2
(45) 17 f 7
(26)
(9) (3) (3) (17)
6)
218
15117 217
3145 (ND)
116
2165
l/81 O/18 l/136 (ND) ’
27155 3119 16123 11/14 24130 100/l 10
J&G4
(29) (25)
(88)
(7)
(16)
(70) (79) (80) (91) 64+ 11
(16)
(49)
(96)
ul
g
&
519
SHORT COMMUNICATIONS
ACKNOWLEDGMENTS We thank Mrs. Annette Pitts, Ms. Shirley Prince and Ms. Rose Kulhavy for excellent technical assistance.
REFERENCES ‘1. 2. 3. 4. 5. 6. 7.
8. 9. 10.
Inman, I. P., and Mestecky, J., Contemp. Top. Mol. Immunol. 3, 111, 1974. Koshland, M. E., Adv. Immunol. 20, 41, 1975. Radl, J., Schuitt, H. R. E., Mestecky, J., and Hijmans, W., Adv. Exp. Med. Biol. 45, 57, 1974. Kaji, H. and Parkhouse, R. M. E., Nature (London) 249,45, 1974. Brandtzaeg, P., and Berdal, P., Stand. J. Immunol. 4, 403, 1975. Sarasombath, S., Mestecky, J., and Skvaril, F., Clin. Exp. Immunol. 29, 67, 1977. Isaacson, P., J. Clin. Pathol. 32, 802, 1979. Mestecky, J., Preud’homme, J.-L., Crago, S. S., Mihaesco, E., Prchal, J. T., and Okos, A. J., Clin. Exp. Immunol. 39, 371, 1980. Yasuda, N., Kanoh, T., and Uchino, H., Clin. Exp. Immunol. 40, 573, 1980. Bast, E. J. E. G., vanCamp, B., Boom, S. E., Jaspers, F. C. A., and Ballieux, R., Clin. Exp. Immunol. 44, 375,
1981.
11. Laurent, Cl., Delsol, Cl., Reyes, F., AbbaI, M., and Mihaesco, E., Clin. Exp. Immunol. 44, 620, 1981. 12. Mason, D. Y., and Stein, H., Clin. Exp. Immunol. 46, 305, 198 1. 13. McCune, J. M., Fu, S. M., and Kunkel, H. G., J. Exp. Med. 154, 138, 1981. 14. Kutteh, W. H., Moldoveanu, Z., Prince, S. J., Kulhavy, R., Alonso, F., and Mestecky, J., Mol. Immunol. 20, 967, 1983. 15. Hajdu, I., Moldoveanu, Z., Cooper, M. D., and Mestecky, J., J. Exp. Med. 158, 1993, 1983. 16. Brandtzaeg, P., Nature (London) 252, 418, 1974. 17. Brandtzaeg, P., Gjeruldsen, S. T., Korsrud, F., Baklien, K., Berdal, P., and Ek, J., J. Immunol. 122, 503, 1979.
18. Korsrud, F. R., and Brandtzaeg, P., Stand. J. 19. Mestecky, J., Winchester, R. J., Hoffman, T., 20. Yasuda, N., Kanoh, T., and Uchino, H., Clin. 21. Reitamo, S., K;iyhkG, K., Konttinen, Y. T.,
Immunol.
13, 281,
1981.
and Kunkel, H. G., J. Exp. Med. Exp. Immunol.
and Bergroth,
46, 142, 198 I. V. J. Histochem.
145, 760, 1977. Cytochem.
31, 843,
1983. 22. Boyurn, A., Stand. J. Clin. Lab. Invest. 21 (Suppl. 97), 77, 1968. 23. Hijmans, W., Schuitt, H. R. E., and Klein, F., Clin. Exp. Immunol. 4, 457, 1969. 24. Crago, S. S., and Mestecky, J., J. Immunol. 122, 906, 1979. 25. Mestecky, J., Zikan, J., Butler, W. T., and Kulhavy, R., Immunochemistry 9, 883, 1972. 26. Kutteh, W. H., Prince, S. J., Phillips, J. O., Spenney, J. G., and Mestecky, J., Gastroenterology
81,
184, 1982. 27. Mayumi, M., Kuritani, T., Kubagawa, H., and Cooper, M., J. Immunol. 130, 671, 1983. 28. Flanagan, J. G., and Rabbitts, T. H., Nature (London) 300, 709, 1982. 29. Crago, S. S., Kutteh, W. H., Moro, I., Allansmith, M. R., Radl, J., Haaijman, J. J., and Mestecky, J., J. Immunol. 132, 16, 1984. 30. Mosmann, T. R., Gravel, Y., Williamson, A. R., and Baumal, R., Eur. J. Immunol. 8, 94, 1978.