Ontogenic development of macrophage subpopulations and

lapositive dendritic cells in fetal and neonatal rat spleen

Motohiro Takeya and Kiyoshi Takahashi
Second Department of Pathology, Kumamoto University School of Medicine, Kumamoto, Japan

Abstract: Development, differentiation, and distribu-

The development of splenic lymphocytes and macro-
tion of macrophage subpopulations and Ia dendritic cells
phages has been studied extensively in animals [4-6] and in
in the fetal and neonatal rat spleen were investigated by
humans [7, 8]. However, information about the fetal and ne-
means of double immunohistochemical staining and im-
onatal development of splenic macrophages is not sufficient,
munoelectron microscopy. To characterize these cell
partly because of the lack of markers for discriminating
populations, a panel of anti-rat macrophage monoclonal
different macrophage subpopulations. The production of
antibodies (RM-1, ED2, ED3, TRPM-3, Ki-M2R) and an
specific monoclonal antibodies against different macrophage
anti-rat Ia antibody (0X6) were used. In the fetal rat
populations has provided an important tool for studying
spleen, macrophages were first detected by RM-1 at fetal
their heterogeneity [9-12]. Using a panel of antimacrophage
day 15. ED2 and/or Ki-M2R macrophages appeared at
antibodies, several investigators have studied the ontogenic
fetal day 16. TRPM-3 and/or ED3 macrophages ap-
development of macrophages in various lymphoid and non-
peared a day later. During the fetal and neonatal develop- lymphoid organs [13-22]. However, the development of mac-
ment, ED2 and TRPM-3 macrophages differentiated in-
rophage subpopulations in the fetal and neonatal spleen is
dependently, maturing into red pulp macrophages and
still not fully understood. In the study reported here, we exa-
marginal metallophilic and marginal zone macrophages mined the ontogenic development of macrophage subpopu-
respectively. Intimate topographical relations were ob- lations in the fetal and neonatal rat spleen using monoclonal
served between ED2 macrophages and hematopoietic
antibodies specific for them.
cells and between TRPM-3 macrophages and marginal
zone lymphocytes. Ia cells were first observed around ar-
terioles at fetal day 15. In the fetal and neonatal period, MATERIALS AND METHODS
the number of Ia cells gradually increased, their shape
became dendritic, and they matured into interdigitating Animals
cells in the inner periarteriolar lymphatic sheath. In on-
togeny, Ia dendritic cells were not stained with ED2 or Wistar white rats maintained under routine laboratory con-
TRPM-3. These results suggest that ED2 macrophages, ditions with free access to food and water were used. The first
TRPM-3 macrophages, and Ia dendritic cells are dis- gestational day (fetal day 0) was decided by the presence of
tinct cell lines that pursue independent developmental a vaginal plug in the morning after mating. Developing
processes in spleen ontogeny. J. Leukoc. Biol. 52: spleens were obtained from rat fetuses at fetal days 14 to 21,
516-523; 1992. from neonates at 0 to 7 days, and from young rats at 10, 14,
17, 21, 24, 28, 35, 42, and 49 days after birth. At least three
Key Words: monoclonal antibodies . immunohistochemistry rat fetuses, newborns, or young adults were examined at
double immunohistochemical staining . immunoelectron microscopy each time point.

Antibodies
INTRODUCTION
Mouse anti-rat macrophage monoclonal antibodies TRPM-3
Macrophages form a heterogeneous cell population with [101 and RM-i [11] were produced in our laboratory. Ki-
regard to their functions, phenotypic expressions, and tissue M2R was kindly provided by Dr. H-H. Wacker, Institute of
localizations. The different organs contain specially differen- Pathology, Kiel University [12]. ED2 and ED3 [9] were pur-
tiated organ-specific macrophages. Because the spleen has a chased from Serotec (Oxford, UK). Monoclonal mouse
unique morphological feature with distinct compartments, anti-rat Ia (OX6, Sera Lab, Sussex, UK) and polyclonal
different macrophage populations reside in different com- rabbit anti-rat immunoglobulin M (IgM)() antibody
partments [1]. Most of the macrophages are located in the ( MBL, Nagoya, Japan) were also used. For immuno-
cord of Billroth in the red pulp (red pulp macrophages). The histochemistry, RM-i, TRPM-3, and Ki-M2R were purified
other macrophages reside in white pulp, which consists of with a protein A-Sepharose affinity column and used at a di-
three major compartments: the periarteriolar lymphatic
sheath (PALS), the lymphoid follicles, and the marginal zone
[2]. Marginal metallophilic macrophages are located in the
periphery of PALS at its border with the marginal zone. Abbreviations: APAAP, alkaline phosphatase anti-alkaline phosphatase;
DAB, 3,3’-diaminobenzidine; 1gM, immunoglobulin M; PALS, periarterio-
Marginal zone macrophages are located in the marginal
lar lymphatic sheath; PBS, phosphate-buffered saline; PLP, periodate-
zone itself. Tingible body macrophages are observed in the lysine-paraformaldehyde.
germinal center of the follicles. In addition, the cells with Reprint requests: Motohiro Takeya, Second Department of Pathology,
dendritic morphology called interdigitating cells are seen in Kumamoto University School of Medicine, 2-2-I Honjo, Kumamoto 860, Japan.
the inner PALS [3]. Received March 9, 1992; accepted July 1, 1992.

516 Journal of Leukocyte Biology Volume 52, November 1992

lution of 1:200. ED2, ED3, and OX6 were used as ascites Immunoelectron microscopy
fluid diluted 1:100. Anti-IgM(j) antibody was used at a dilu-
Selected splenic tissues were sliced with razor blades and
tion of 1:200.
fixed in 4% PLP at 4#{176}C
for 4 h. After washing with PBS, the
slices were incubated with TRPM-3 or ED2 overnight at
Immunohistochemistry 4#{176}C. Then they were reacted with peroxidase-labeled
anti-mouse immunoglobulin [F(ab’)2] for 3 h at room tempera-
Immediately after resection, the splenic tissues were fixed in
ture. Peroxidase activity was visualized as described above.
2% periodate-lysine-paraformaldehyde (PLP) fixative for 4 h
Then the tissues were postfixed with 1% osmium tetroxide in
at 4#{176}C.After 4-h washes with phosphate-buffered saline
0.05 M cacodylate buffer (pH 7.4), dehydrated in a graded
( PBS) containing 10, 15, or 20% sucrose and 20% sucrose
series of ethanols, and embedded flat in epoxy resin. Ultra-
+ 5% glycerol, they were embedded in tissue-embedding
thin sections were cut on an Ultrotome Nova (LKB, Up-
medium (Miles, Elkhart, IN), frozen in dry ice-acetone, and
psala, Sweden) and observed in a Hitachi-12A electron
cut into 6-sm-thick sections with a cryostat (Bright, Hun-
microscope (Hitachi, Tokyo, Japan) after staining with ura-
tingdon, UK). After inhibition of endogenous peroxidase ac-
nyl acetate.
tivity according to the method of Isobe et al. [23], the sec-
tions were incubated with each monoclonal or polyclonal
antibody for 60 mm at room temperature. After three washes RESULTS
with PBS for 5 mm each, peroxidase-conjugated sheep
anti-mouse immunoglobulin [F(ab’)2] diluted 1:100 (Amer- Localization of macrophage subpopulations in the adult
sham, Amersham, UK) was reacted for 60 mm as the second rat spleen
step. To visualize peroxidase activity, 3,3’-diaminobenzidine
Table 1 summarizes the positive cells of each antibody in the
(DAB) (Sigma, St. Louis, MO) was used as substrate in 0.05
adult rat spleen. The reactivity of each antimacrophage
M Tris-HC1 buffer (pH 7.6) containing 0.01% H202.
monoclonal antibody in the adult spleen is almost the same
Nuclear stain was performed with hematoxylin. Control
as those described in the previous reports [9-12]. RM-1 was
slides were incubated with nonimmunized mouse serum or
reactive with most splenic macrophages and interdigitating
PBS instead of the primary antibody; they were invariably
cells (data not shown). ED2, Ki-M2R, ED3, and TRPM-3
negative.
recognized different macrophage subpopulations. The
former two were reactive with red pulp macrophages,
Double immunohistochemical staining whereas the latter two labeled marginal metallophilic macro-
phages and marginal zone macrophages. Subset specificities
As the first step, the frozen sections were stained by the in- of these antibodies were well documented by double im-
direct immunoperoxidase method mentioned above using munohistochemical staining. In the double staining of ED2
one of the monoclonal antibodies. After visualizing this reac- and TRPM-3 (Fig. la), ED2 recognized red pulp macro-
tion stain brown by DAB, the sections were treated with 0.1 phages exclusively, whereas TRPM-3 labeled marginal
M glycine-HC1 buffer (pH 2.2) for 60 mm with four changes
metallophilic macrophages and marginal zone macrophages.
to remove the first and second antibodies reacted. Then the Interdigitating cells in the inner PALS and B cells in the
same sections were incubated overnight with one of the other marginal zone and lymphoid follicles were detectable by
antimacrophage monoclonal antibodies. After three washes their expression of Ia antigen (Fig. ib) as reported previously
with Tris-buffered saline (pH 7.6), the sections were treated [5, 6, 24]. B lymphocytes in the marginal zone were strongly
by the alkaline phosphatase anti-alkaline phosphatase positive for 1gM, and those in the follicles were moderately
(APAAP) method using an APAAP kit (DAKO, Carpinteria,
positive for 1gM (data not shown).
CA). To visualize the reaction of APAAP, the sections were
In the immunoelectron microscopic study, ED2 distinctly
stained blue in a substrate made up of0.2 mM naphthol AS-
labeled the cell membrane of red pulp macrophages (Fig.
MX phosphate (Sigma), 1 mM fast blue BB salt (Sigma),
2a). TRPM-3 clearly labeled the cell membrane of marginal
and 1 mM levamisole (Aldrich, Milwaukee, WI) in 50 mM
metallophilic macrophages, which formed a rim surrounding
Tris-HC1 buffer (pH 8.5) for 20 mm. To confirm the reaction
the PALS (Fig. 2b).
specificities of each step, the first antibodies in the first or se-
cond step were omitted in control experiments.
Development and difterentiation of macrophage
Double staining of rabbit anti-rat IgM() polyclonal anti-
subpopulations in the fetal rat spleen
body and one of the antimacrophage monoclonal antibodies
was performed as above without the treatment with glycine- At fetal day 14, the spleen was recognizable as the red streak
HC1 buffer. attached to the dorsal wall of the stomach. At this stage small

TABLE 1. Reactivity of Antirat Antibodies in Adult Rat Spleen

Antibody Isotype Site of recognition Reactive cells

RM-1 IgGl Cell membrane Monocytes, most macrophages, interdigitating cells

ED2 IgG2a Cell membrane Red pulp macrophages
Ki-M2R IgA Cell membrane Red pulp macrophages
ED3 IgG2a Cell membrane Marginal metallophilic macrophages, marginal zone macrophages
TRPM-3 IgG2a Cell membrane Marginal metallophilic macrophages, marginal zone macrophages
0X6 IgGl Cell membrane Interdigitating cells, B lymphocytes
anti-IgM(gz) Polyclonal Cytoplasm, cell membrane B lymphocytes in follicles and marginal zone

Takeya and Takahashi Development of rat splenic macrophages 517

Fig. 1. Immunohistochemical staining of adult rat spleen. A, arteriole; F, lymphatic follicle; MZ, marginal zone; P, periarteriolar lymphatic sheath; RP,
red pulp. (a) In double staining, ED2 (brown) specifically labels red pulp macrophages, whereas TRPM-3 (blue) stains marginal metallophilic macrophages
and marginal zone macrophages. x 100. (b) OX6 (anti-Ia) is reactive with interdigitating cells in inner PALS and B cells in the marginal zone and lymphoid
follicles. xlOO.

clusters of erythroblasts were scattered between mesen- were seen farther away (Fig. 3g). Ia dendritic cells were situ-
chymal cells. At fetal day 15, a few RM-1 macrophages ap- ated mainly around the arterioles and their number gradu-
peared dispersively. Some Ia cells were also seen around the ally increased. IgM cells were not observed in the fetal
arterioles (Fig. 3a). Although these cells were round or oval, spleen.
they were thought to be the precursor cells of interdigitating
cells, not of B cells, because they were also labeled by RM-1
Development and differentiation of macrophage
in double staining (Fig. 3b). At fetal day 16, a few ED2 (Fig. subpopulations in the spleen of rat neonates and young
3c) and Ki-M2R cells appeared. TRPM-3 (Fig. 3d) and rat adults
ED3 cells first appeared at fetal day 17. Through the fetal In the very early neonatal period, from day 0 to 5, many Ia
stage, RM-P macrophages outnumbered ED2 macro- cells with short and blunt cell surface projections accumu-
phages (Fig. 3e) and TRPM-3 macrophages (Fig. 30. In lated around the arterioles (Fig. 3h). ED2 (and Ki-M2R)
double staining, most ED2 macrophages (Fig. 3e) and macrophages and TRPM-3 (and ED3) macrophages were
TRPM-3 macrophages (Fig. 31) were also labeled by RM-1, scattered outside the Ia dendritic cells. In double staining,
whereas ED2 and TRPM-3 labeled different macrophage Ia cells were not reactive with ED2 or TRPM-3 (Fig. 3i).
populations (Fig. 3g). In contrast to the adult spleen, ED2 At day 2, IgM B lymphocytes were first observed around the
macrophages and TRPM-3 macrophages were intermixed. arterioles, and their number increased thereafter to form
However, TRPM-3 macrophages tended to accumulate primitive marginal zone. At day 5, ED2 macrophages were
closely around the arterioles, whereas ED2 macrophages found outside the primitive marginal zone (Fig. 3j), whereas

Fig. 2. Immunoelectron micrographs of adult rat spleen. (a) Cell membrane of a red pulp macrophage (M) is labeled by ED2. (b) Reaction products of
TRPM-3 are observed on the cell membrane of marginal metallophilic macrophages (M). Bar, I zm.

518 Journal of Leukocyte Biology Volume 52, November 1992

Takeya and Takahashi Development of rat splenic macrophages 519
 Fig. 3. Immunostaining of fetal and neonatal rat spleen. A, arteriole; MZ, marginal zone; P, periarteriolar lymphatic sheath; RP, red pulp. (a) A few Ia”
cells are seen around an arteriole at fetal day 15. x520. (b) Double staining with Ia (brown) and RM-l (blue) at fetal day 15. Most La’ cells (arrowheads)
are labeled by RM-l, whereas some RM-l” cells (arrows) are devoid of staining with Ia. x800. (c) ED2 macrophages (arrowheads) first appeared in fetal
spleen at day 16. x 520. (d) TRPM-3 macrophages (arrowheads) first appeared in fetal spleen at day 17. x 520. (e) Double staining with ED2 (brown)
and RM-l (blue) at fetal day 17. Arrowheads indicate ED2 and RM-l” macrophages. x400. (I) Double staining with TRPM-3 (brown) and RM-l (blue)
at fetal day 18. Arrowheads indicate TRPM-3” and RM-l” macrophages. x400. (g) In double staining of a fetal spleen at day 20, ED2 (brown) and
TRPM-3 (blue) recognize different macrophage populations. x 260. (h) Ia” cells are mainly observed around an arteriole at I day after birth. x 260. (i)
Double staining of a neonatal spleen at 1 day after birth. TRPM-3” cells (brown) are scattered outside the Ia” cells (blue). x 400. (j) At neonatal day 5,
ED2 (blue) macrophages were found outside the primitive marginal zone, which is formed by 1gM” B lymphocytes (brown). x200. (k) ED2 (brown)
macrophages are seen outside the rim of TRPM-3” marginal metallophilic macrophages (blue) at neonatal day 5. x 260.

TRPM-3 marginal metallophilic macrophages formed a loped, ED2 and TRPM-3 labeled different macrophage sub-
rim surrounding the primitive PALS, although the cells were populations. Thus, it is considered that ED2 and TRPM-3
not continuously arrayed (Fig. 3k). macrophage subpopulations differentiate independently
On immunoelectron microscopy of early neonatal spleen, through the fetal and neonatal life and mature into red pulp
TRPM 3 macrophages extended cytoplasmic processes macrophages and marginal metallophilic and marginal zone
among the surrounding lymphocytes (Fig. 4a). ED2 macro- macrophages, respectively. The difference between these two
phages, on the other hand, were usually found between the major macrophage subpopulations was also observed in an
hematopoietic cells (Fig. 4b). After day 5, the PALS gradu- experiment on liposome-mediated elimination of splenic
ally expanded. At day 10, TRPM-3 marginal metallophilic macrophages by van Rooijen et al. [26], who reported that
macrophages formed a complete rim between the PALS and red pulp macrophages and marginal zone macrophages
the marginal zone, and a few TRPM-3 marginal zone mac- showed a striking difference in the kinetics of their reappear-
rophages appeared among IgM marginal zone B lympho- ance. Considering their result, our data indicate that these
cytes (Fig. Sa). The marginal zone bridging channels [25] two major splenic macrophages are distinct cell lineages.
were also recognized at this stage (Fig. 5a). At day 14, IgM
and Ia B lymphocytes accumulated inside the rim of
TRPM-3 marginal metallophilic macrophages to form
primary lymphoid follicles. At the same day, many Ia inter-
digitating cells with elongated cell processes were observed in
the inner PALS (Fig. Sb). At day 28, germinal centers deve-
loped in the lymphoid follicles as reported previously [4].
Thus the architecture of the white pulp became very similar
to that of the adult spleen. Figure 6 schematically summa-
rizes the development ofmacrophage subpopulations and Ia
dendritic cells in the rat spleen.

DISCUSSION

Immunohistochemistry with monoclonal antibodies readily

permits the differentiation of various cell types. In recent
years, many antimacrophage monoclonal antibodies have
been used to investigate macrophage heterogeneity. Using a
number of monoclonal antibodies, Buckley et al. [1] demon-
strated in the human spleen that different phenotypic subsets
of macrophages are present in discrete microanatomic loca-
tions. Among the antirat monoclonal antibodies used in this
study, RM-1 is considered to be an anti-pan-macrophage
monoclonal antibody that recognizes almost all macro-
phages, monocytes, and dendritic cells such as interdigitat
ing cells or Langerhans cells [11]. ED2 and Ki-M2R recog-
nize resident macrophages in various organs but not blood
monocytes. In the spleen, they react exclusively with red
pulp macrophages [9, 12]. TRPM-3 recognizes certain mac-
rophage subpopulations, such as marginal metallophilic
macrophages and marginal zone macrophages in the spleen,
sinus macrophages in the lymph nodes, and omentum mac-
rophages, but not blood monocytes [10]. ED3 revealed a
reaction pattern similar to that of TRPM-3 [9]. Because
ED2 and TRPM-3 gave a clear reaction to the cytoplasmic
membrane among these macrophage-specific antibodies,
Fig. 4.
Immunoelectron micrograph of neonatal spleen at day 3. (a) A
these two antibodies were mainly used for the immunoelec-
TRPM-3” macrophage (M) extends its cytoplasmic processes between sur-
tron microscopic study. rounding lymphocytes. Distinct reaction products (arrowheads) are seen on
It is noteworthy that the different macrophage populations the whole plasma membrane of the macrophage. (b) An ED2 macrophage
express TRPM-3 and ED2 through their development. Even (M) shows intimate cell-to-cell contact with surrounding erythroblasts. Ar-
at the fetal period, when no splenic compartment is deve- rowheads indicate reaction products of ED2. Bar, I gzm.

520 Journal of Leukocyte Biology Volume 52, November 1992

Fig. 5. Immunostaining of young rat spleen. A, arteriole; BC, marginal zone bridging channel; F, lymphoid follicle; MZ, marginal zone; P periarteriolar
lymphatic RP, red pulp.
sheath; (a) Double staining with 1gM (brown) and TRPM-3 (blue) at day 10. TRPM-3 marginal metallophilic macrophages (blue)
form a complete rim between PALS and marginal zone. A marginal zone bridging channel is also seen. x 130. (b) Ia dendritic cells are seen in the inner
PALS. Lymphocytes in marginal zone and a primary lymphoid follicle are also positive for Ia. x 100.

The marginal zone is the largest white pulp compartment arterioles in the fetal period before the emigration of IgM
in the rat [27] and has a unique property both morphologi- marginal zone B cells. After the appearance of marginal
cally and functionally. Marginal zone B cells are considered zone B cells shortly after birth, an intimate topographical re-
to be a distinct cell lineage and are easily detectable by their lation between TRPM-3 macrophages and marginal zone B
expression of IgM() [28]. Marginal zone macrophages play cells was observed by immunoelectron microscopy. Taking
a special role in presenting thymus-independent antigens to these findings together, TRPM-3 macrophages are consi-
surrounding B lymphocytes [29]. In the present study, dered to participate in the emigration and differentiation
TRPM-3 macrophages were already observed around the processes of marginal zone B cells.

A#{149}PA 4’
V#{174}
*
:4f N
)::: ts;s5;::c )RP

fetal 20 d 5d lOd

ladendritic cell
-- ED2 macrophage
TRPM4’ macrophage
14d 28d
Fig. 6. Schematic drawing of the ontogenic development of ED2 and TRPM-3” macrophage subpopulations and Ia dendritic cells in rat spleen. A, arteri-
ole; BC, marginal zone bridging channel; PF, primary lymphoid follicle; GC, germinal center; MZ, marginal zone; P, periarteriolar lymphatic sheath;
RP, red pulp.

Takeya and Takahashz Development of rat splenic macrophages 521

 In rodents such as rats and mice, the spleen, particularly monoclonal antibodies ED1, ED2 and ED3. Immunology 54,
the red pulp, is a major site of hematopoietic activity in the 589-599.
10. Takeya, M., Hsiao, L., Takahashi, K. (1987) A new monoclonal
fetal and neonatal life, and splenic hematopoiesis persists un-
antibody, TRPM-3, binds specifically to certain rat macro-
til adulthood [30]. As ED2 macrophages were observed at
phage populations: immunohistochemical and immunoelectron
the center of the erythroblastic islands, these cells may play
microscopic analysis. j Leukoc. Biol. 41, 187-195.
an important role in fetal and neonatal erythropoiesis. Con- 11. Takeya, M., Hsiao, L., Shimokawa, Y., Takahashi, K. (1989)
sidering the different microanatomical localizations of ED2 Heterogeneity of rat macrophages recognized by monoclonal
and TRPM-3 macrophages, it is of great interest to note antibodies: an immunohistochemical and immunoelectron
that these two macrophage subpopulations share special microscopic study. J. Histochem. Cytochem. 37, 635-641.
microenvironments related to their functions at the early ne- 12. Wacker, H-H., Razdum, H.J., Parwarewsch, R. (1985) Ki-M2R,
onatal period. a new specific monoclonal antibody, discriminates tissue macro-
phages from reticulum cells and monocytes in vivo and in vitro.
In ontogeny, the thymus is the initial organ for the expres-
j Leukoc. Biol. 38, 509-520.
sion of Ia antigens [31-33]. In the rat thymus, we observed
13. Dijkstra, CD., van Rees, E.P., D#{246}pp, E.A. (1988) Ontogeny of
that Ia primitive/fetal macrophages appeared in the thymic
macrophages and dendritic cells of the rat. Adv. Exp. Med. Biol.
primordium at fetal day 14 and differentiated into inter-
237, 731-736.
digitating cells thereafter[14]. In the present study of the rat 14. Hsiao, L., Takahashi, K., Takeya, M., Arao, T. (1990) Differen-
spleen, Ia cells were first observed around the arterioles at tiation and maturation of macrophages into interdigitating cells
fetal day 15. Thereafter, more Ia cells accumulated around and their multicellular complex formation in the fetal and post-
the arterioles and developed into dendritic cells at the same natal rat thymus. Thymus 17, 219-235.
site. Barclay [24] demonstrated that Ia cells in the inner 15. Hsiao, L., Takeya, M., Arao, T., Takahashi, K. (1989) An im-
PALS were interdigitating cells in the adult rat spleen. As munohistochemical and immunoelectron microscopic study of
the ontogeny of rat Langerhans cell lineage with anti-rat macro-
these Ia dendritic cells were also labeled by RM-1, they were
phage and anti-Ia monoclonal antibodies. j Invest. Dermatol. 93,
thought to be precursor cells of interdigitating cells but not
780-786.
B lymphocytes. Throughout the fetal and neonatal period,
16. Izumi, S., Takeya, M., Takagi, K., Takahashi, K. (1990) Onto-
Ia dendritic cells were not labeled by ED2 or TRPM-3 in genic development of synovial A cells in fetal and neonatal rat
the double immunohistochemical staining. These data mdi- knee joints. Cell Tissue Res. 262, 1-8.
cate that Ia dendritic cells form a distinct cell lineage with 17. Morris, L., Graham, CF., Gordon, S. (1991) Macrophages in
an independent differentiation process. haemopoietic and other tissues of the developing mouse de-
In summary, it is considered that Ia dendritic cells, red tected by monoclonal antibody F4/80. Development 112, 517-526.
pulp macrophages, and marginal metallophilic and marginal 18. Naito, M., Takahashi, K., Nishikawa, S. (1990) Development,
differentiation, and maturation of macrophages in the fetal
zone macrophages pursue different emigratory and differen-
mouse liver. j Leukoc. Biol. 48, 27-37.
tiation processes in the fetal and neonatal rat spleen, as
19. van Rees, ER, van der Ende, MB., Sminia, T. (1991) Ontogeny
shown schematically in Fig. 6. of macrophage subpopulations and Ia-positive dendritic cells in
pulmonary tissue of the rat. Cell Tissue Res. 263, 367-373.
20. Takahashi, K., Naito, M., Katabuchi, H., Higashi, K. (1991)
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