Macrophages of human first trimester decidua express markers associated to alternative activation.

PROBLEM: Depending on the type of their activation, macrophages may promote TH1- or TH2-type of immune responses. To date, not much is known about the activation phenotype of decidua macrophages, which - together with NK cells - constitute the majority of bone marrow derived cells at this location. METHOD OF STUDY: The study was based on analysis of healthy first trimester decidua by immunohistochemistry and flow cytometry. We analyzed expression of markers characteristic for alternatively activated macrophages (Mphi2). RESULTS: The markers MS-1 (stabilin-1) and coagulation factor XIIIa were found expressed in the interior of decidua macrophages (DMphi). In contrast, indoleamine 2,3-dioxygenase (IDO), an enzyme induced in macrophages by IFNgamma, was not present in DMphi. CONCLUSIONS: First trimester DMphi display phenotypic markers associated to alternatively activated macrophages. In addition, absence of IDO indicates that DMphi are not under a predominant influence of IFNgamma.

B7 family molecules are favorably positioned at the human maternal-fetal interface.

The human placenta utilizes both active and passive mechanisms to evade rejection by the maternal immune system. We investigated the pattern of expression of the B7 family of immunomodulatory molecules B7-H1 (PD-L1), B7-2 (CD86), and B7-1 (CD80) at the term maternal-fetal interface. Northern blot and reverse transcription-polymerase chain reaction (RT-PCR) analyses showed that B7-H1 mRNA is abundant in term placenta and that cytotrophoblasts are sources of this message. Immunohistochemistry demonstrated that B7-H1 is constitutively expressed by the syncytiotrophoblast and by extravillous cytotrophoblasts, both of which are juxtaposed to maternal blood and tissue. By contrast, placental stromal cells, including macrophages, lacked the protein. Expression of B7-H1 protein was low in first-trimester placenta compared to second- and third-trimester tissue (P < 0.05) and was enhanced in cultured cytotrophoblasts by treatment with either interferon-gamma or epidermal growth factor (P < 0.05), suggesting that one or both of these mediators regulates B7-H1 expression in the placenta. RT-PCR and immunofluorescence analysis of term placental tissue revealed different patterns of expression of the immunostimulatory protein, B7-2. In contrast to B7-H1, B7-2 mRNA and protein were absent in cytotrophoblast cells but present in maternal macrophages and some fetal macrophages. The B7-1 mRNA and protein were absent at the maternal-fetal interface. These studies document expression of the B7 family proteins at the maternal-fetal interface and demonstrate that B7-H1 is positioned such that it could facilitate protection of fetal cells against activated maternal leukocytes. Conversely, B7-2 was absent on trophoblasts and was appropriately localized to fetal and maternal macrophages, which may participate in antigen presentation.

Absence of HLA-G expression in macrophages of human decidua.

PROBLEM: Macrophages - together with natural killer (NK) cells - constitute the majority of bone marrow derived infiltrating cells in the decidua. As interferon-gamma (IFN-gamma), a cytokine produced by NK cells, has been reported to induce expression of human leukocyte antigen (HLA-G) in monocytic cells, suggesting expression of HLA-G on decidua macrophages potentially stimulated by IFN-gamma, the question arises whether decidua macrophages in normal pregnancy express HLA-G. METHOD OF STUDY: The study was based on immunohistochemistry and flow cytometry. In order to exclude that potentially elusive soluble HLA-G was not detected by immunohistochemistry, we performed in addition RT-PCR of flow-sorted decidua macrophages. RESULTS: Our findings indicate that HLA-G is not present on macrophages of first trimester or term decidua in either membrane-bound or soluble form. Transcripts for soluble HLA-G1 and -G2 were not detected. CONCLUSIONS: We exclude a role of HLA-G on the surface of decidua macrophages or of soluble HLA-G1 or -G2 as a secretory product of decidua macrophages with regard to interaction with HLA-G receptors present in or outside the decidua.

Decidual macrophages are potentially susceptible to inhibition by class Ia and class Ib HLA molecules.

Several members of the immunoglobulin-like transcript (ILT), also called leukocyte immunoglobulin-like receptor (LIR), family of transmembrane proteins have been identified as receptors for class I HLA molecules and transduce inhibitory signals to leukocytes upon binding of these ligands. The ligands for ILT2 (LIR1/CD85j) and ILT4 (LIR2/CD85d) include HLA-A, -B, and -G, the last of which is highly expressed in fetal trophoblast cells in both membrane-bound and soluble isoforms. To investigate the potential of fetally-derived HLA class I molecules to interact with maternal macrophages through these receptors, we examined the expression patterns of ILT2 and ILT4 in decidual macrophages. Highly purified populations of decidual macrophages were obtained by fluorescence activated cell sorting and were examined by RT-PCR for these messages. Analysis of mRNA from first trimester and term macrophages, as well as the monocyte cell line U937, resulted in amplicons of similar size to those expected for ILT2 and ILT4. Sequence analysis of the amplicons revealed that the messages from decidual macrophages corresponded to ILT2 and ILT4 messages. The message amplified from the U937 cells using the ILT2 primers was also found to be identical to ILT2; however, sequence analysis revealed that the ILT4 message amplified from these cells is a truncated form of the message. Dual label flow cytometry confirmed the expression of ILT2 and ILT4 on CD14-positive first trimester decidual macrophages and U937 cells. These results reveal that inhibitory HLA receptors are expressed in decidual macrophages and suggest that HLA-G may deliver negative signals to maternal decidual macrophages through interaction with these receptors.