Activation of resting human B cells by helper T-cell clone supernatant: characterization of a human B-cell-activating factor.

The effects of helper T-cell clone supernatants on resting human B cells were investigated. Four different helper T-cell clones (two T4+ and two T8+) were stimulated by anti-T3 monoclonal antibodies on Sepharose beads or anti-T11(2) plus anti-T11(3) monoclonal antibodies. The supernatants from these activated clones induced the proliferation of highly purified resting B lymphocytes from the peripheral blood. The B cells exhibited a cell size and a surface-antigen pattern (4F2 antigen and transferrin receptor) of phase G0 B cells, and they were functionally resting. In response to T-cell supernatants a large fraction of the B cells enlarged and expressed 4F2 antigens and transferrin receptors. In gel filtration, the corresponding activity migrated with an apparent Mr of 12,000-15,000. Our findings strongly support the existence of a human B-cell-activating factor acting on resting B cells and causing them to enter phase G1 of the cell cycle.     

Transfected rat islet tumour cells express mouse major histocompatibility complex class I antigens functionally

Summary Rat insulinoma cells clone 5AH-B, were transfected by electroporation with the gene encoding the mouse major histocompatibility complex class I antigen H-2K^b, whereupon stable transfectants were selected and analysed. Data from flow cytometric analyses using three different H-2K^b specific monoclonal antibodies and functional assays using H-2K^b specific alloimmune cytotoxic T cells revealed that the encoded H-2 antigen was expressed in a functional manner. Similar experiments employing the monoclonal antibody OX-18, which recognizes rat major histocompatibility class I molecules, and xenoimmune cytotoxic T cells specific for the endogenously expressed RT1^g antigen showed that functional expression of the RT1^g antigen was maintained. However, a down-regulation of the expression was observed in H-2K^b positive transfectants, whereas normal expression was retained in K^b negative transfectants. The function of the native promoters of both the endogenous and the transfected class I genes was found to be preserved in the transfectants as assessed by the response to stimulation with interferon-. The present study was unable to confirm the reports of RIN specific lysis by T cells from multiple low dose streptozotocin diabetic mice. Even in the presence of the syngeneic restriction element no lysis was observed. We conclude, that rat insulinoma cells clone 5AH-B, are able to integrate a foreign class I antigen gene and express the encoded product functionally. The data also suggest the possibility of creating major histocompatibility antigen positive rat insulinoma cells which are RT1^g negative. Such transfectants will be of great potential value for the dissection of cell mediated B cell destructive processes.     

Major histocompatibility complex-linked immune-responsiveness is acquired by lymphocytes of low-responder mice differentiating in thymus of high-responder mice.

Female murine T cells can respond to the Y antigen of male cells by generating cytotoxic T-killer lymphocytes. Responsiveness is linked to several H-2 genes. Two types of low responders can be distinguished: the B10.A(5R) (H-2i5) strain, a low responder because it lacks Y-specific precursor T cells able to differentiate into cytotoxic T-killer cells; and the CBA/J (H-2k) strain, a low responder because it lacks Y-specific T-helper cells able to support differentiation of T-killer cell precursors. B10.A(5R) stem cells differentiating in an x-irradiated (CBA/J X C57BL/6) (H-2k X H-2b)F1 host respond to Y antigen by generating T-killer cells whereas CBA/J stem cells do not. The results are consistent with the hypothesis that diversity of T-cell receptors is generated by somatic mutation of germ-line genes encoding specificity for self-H-2. A detailed account of this hypothesis is presented.     

Major histocompatibility complex class I-restricted activation of cloned T cells by a soluble protein in the absence of accessory cells

A T-cell clone, 10BK.1, was established from the draining lymph nodes of (B10 x B10.BR)F1 mice immunized with ovalbumin (OVA) according to standard protocols. Upon coculture with the antigen, 10BK.1 cells reacted by production of lymphokines and by proliferation despite the absence of additional antigen-presenting cells. These T cells do not express major histocompatibility complex (MHC) class II molecules on the cell surface as assessed on the basis of several criteria: by cytofluorometric analysis I-A and I-E determinants were not detectable; 10BK.1 cells could not act as antigen-presenting cells for long-term-cultured MHC class II-restricted T-cell clones; and monoclonal antibodies directed at both MHC class II isotypic complexes (I-A, I-E) did not suppress their OVA-induced activation. In contrast, proliferation of 10BK.1 T cells in response to OVA was abrogated by antibodies directed at H-2Kb antigens. Inhibition experiments employing antibodies directed at Lyt-2 and L3T4 antigens in addition to cytofluorometric analysis revealed that T-cell clone 10BK.1 exhibits the Thy-1+,Lyt-2+,Ly-1-,L3T4- phenotype. 10BK.1 cells pulsed with OVA and fixed with glutaraldehyde induced proliferation of untreated 10BK.1 cells. These data support the theory that 10BK.1 T cells present the exogenous globular protein OVA to one another in an MHC class I-restricted manner, resulting in cell activation and proliferation independent of added accessory cells.     

T-cell antigen receptors with identical variable regions but different diversity and joining region gene segments have distinct specificities but cross-reactive idiotypes.

The T-cell antigen receptor alpha-chain genes of an alloreactive, H-2Db-specific cytotoxic T-cell clone (3F9) are described. This study and our work on the 3F9 beta-chain genes reveal that the variable region gene segments for the alpha and beta chains expressed in 3F9 are identical to the ones used by a chicken erythrocyte-specific, I-Ab-restricted helper T-cell clone (LB2). These two clones differ, however, in the diversity and joining portions of the alpha and beta chains of their T-cell receptor molecules. The analysis of 3F9 and LB2 with monoclonal antibodies specific for the 3F9 T-cell receptor shows that these two T-cell clones share the same idiotype; however, 3F9 and LB2 do not exhibit any antigen and/or major histocompatibility complex cross-reactivity. This suggests that the diversity and joining regions of the T-cell receptor may play a key role in antigen and/or major histocompatibility complex recognition.     

Absence of H-2 antigens capable of reacting with cytotoxic T cells on a teratoma line expressing a T/t locus antigen.

An established cell line of murine teratoma cells (F9), which lacks serologically detectable H-2 that is determined by a wild-type T/t locus gene. These cells are not killed and do not react with cytotoxic T cells sensitized to H-2 antigens in a cell-mediated lympholysis assay. Modification of spleen cells from the strain or origin (129) of this teratoma line with trinitrobenzene sulfonic acid allows the generation of syngeneic killer cells that display a cytotoxic effect against trinitrophenyl-modified splenic targets, but not against trinitrophenyl-modified F9 targets. Thus, the F9 antigen is structurally similar to H-2b but does not act as a target antigen in the cell-mediated lympholysis assay for anti-H-2b cytotoxic T cells, nor does it crossreact with H-2b antigens at the T cell level.     

Monoclonal antibodies to thyroglobulin elucidate differences in protein structure of thyroglobulin in healthy individuals and those with papillary adenocarcinoma

Monoclonal antibodies specific for human thyroglobulin (Tg) from normal subjects were prepared by the hybridoma technique. Antibodies from three clones (clones B2F, C6E, and C6G) were found to produce linear Scatchard plots, as predicted for homogeneous antibodies. Based on different patterns of cross-reactivity with Tg from various species, these monoclonal antibodies recognized different determinants on the Tg molecule. Moreover, antibodies from clone B2F bound simultaneously with clone C6E or C6G to Tg. Therefore, antibody from clone B2F must bind to a site on Tg distant from those recognized by clone C6E or C6G. The monoclonal antibodies C6G and C6E bound almost equally to normal Tg and Tg from patients with Graves' disease, adenoma, follicular carcinoma, and papillary adenocarcinoma. In contrast, whereas clone B2F bound equally well to normal Tg and Tg from patients with Graves' disease, adenoma, and follicular carcinoma, this clone bound poorly to Tg from patients with papillary adenocarcinoma. Since the binding activity of clone B2F for unfolded or degenerated Tg was remarkably decreased, these differences in binding activities to native Tg may reflect changes in conformation of the Tg molecule. Thus, the results indicate there may be conformational changes in Tg from patients with different thyroid diseases.     

Anti-L3T4 antibody inhibits the lysis of H-2 class II antigen-negative target cells by L3T4+ cytotoxic T lymphocytes

Anti-L3T4 monoclonal antibodies inhibit the cytotoxic activity of L3T4+ cytotoxic T lymphocytes specific for H-2 class I antigens. The P815 target cells used to detect this population of murine cytolytic cells are shown by immunofluorescence, radioimmunoprecipitation, and RNA blot analysis not to express H-2 class II protein or mRNA. Contrary to previously proposed models regarding its function, we conclude that the L3T4 molecule is involved at some stage of the lytic interaction between the class I-specific L3T4+ effector cell and its target cell by a mechanism for which there is not an obligatory requirement for H-2 class II antigen expression by the target cell. L3T4 may be an early component of the system that transduces the activation signal from the T-cell receptor complex to the cytoplasm, a cell-surface receptor for a yet undefined natural ligand that delivers a negative signal to the killer T cell, or it may modulate the avidity of the antigen-specific T-cell receptor through a direct physical association with it.     

乙型肝炎表面抗原与粒细胞巨噬细胞集落刺激因子融合表达质粒诱导小鼠特异性免疫应答

目的 观察乙型肝炎表面抗原(HBsAg)与粒细胞巨噬细胞集落刺激因子(GM CSF)融合表达质粒诱导特异性免疫应答的效果。方法 以HBsAg与GM-CSF融合表达质粒DNA免疫BALB/C小鼠,用酶链免疫吸附方法检测质粒诱导BALB/C小鼠产生抗HBs及脾细胞经HBsAg体外刺激后分泌白细胞介素(IL)-2、4及γ干扰素(IFNγ)的水平;并用~(21)Cr释放法检测HHsAg特异性细胞毒性T淋巴细胞(CTL)反应。结果 各组小鼠加强免疫后血清抗体水平(P/N值)A组(PcDNA3.1-S)16.1 20.3,B组(PcDNA3.1 GM-CSF-S)28.3±19.7,F组(重组酵母乙型肝炎疫苗)29.5±21.5,C组(PcDNA3.1S-GM-CSF)0.3±0.0,D组(PcDNA3.1—S PcDNA3.1-GM-CSF)4.5 5.9,E组(PcDNA3.1—GM-CSF)0.9±1.2,G组(PcDNA3.1)及H组(PBS)为阴性(F-4.176,P<0.01);IL-2分泌水平A组(26.6±1.9)、B组(56.0±2.2)、C组(4.3±1.2)、D组(33.5±1.7)、F组(3.5 1.4)、F组(7.5±2.0)、G组(1.5±0.7),H组未检测到IL-2分泌(F=31.188,P<0.01);IFN-γ分泌水平A组(64.0±7.5)、B组(139.0±17.0)、C组(11.0±5.0)、D组(53.0±9.5)、E组(9.0±3.0)、F组(13.0±2.0)、G组(8.0±2.0,P(0.05),H组未检测到IFN—γ泌(F=31.796,P<0.01);HBsAg特异性CTL活性,效靶比为30:1及10:1时各组音有差异有显著性(F值分别为26.891、12     

Attachment of an anti-receptor antibody to non-target cells renders them susceptible to lysis by a clone of cytotoxic T lymphocytes.

The molecular basis for the dependence of antigen recognition by T cells on products of the major histocompatibility complex (MHC) is unknown, and the antigenic structures that are actually bound by T-cell receptors are ill-defined. In this study, we asked whether a monoclonal antibody (mAb) that reacts with the T-cell receptor of a clone of murine cytotoxic T lymphocytes (CTL) and not with the receptors of other CTL clones can substitute for that clone's natural ligand in specific cytolytic reactions. To answer the question, a mAb (1B2) to the receptor of a CTL clone (2C) was attached covalently to 51Cr-labeled cells that were not otherwise susceptible to lysis by clone 2C, and the cells thus modified were then tested as targets for clone 2C and other CTL clones of similar specificity. All labeled cells modified in this way, including a murine cell line that expresses no cell-surface MHC class I molecules and a human cell line, were lysed by clone 2C but not by other CTL clones. If, however, instead of attaching the mAb to the receptor of clone 2C, the cells were modified by attaching to them mAbs to other surface antigens on CTL [lymphocyte function-associated antigen (LFA-1), Thy-1.2], they were not lysed. In cytolytic titrations, the cells that had been converted by attachment of mAb 1B2 into specific targets for clone 2C were just as susceptible to lysis by that clone as the clone's natural H-2d targets (e.g., P815 cells). However, some accessory surface molecules (LFA-1, Lyt-2) that are required for clone 2C to lyse its natural H-2d targets seemed not to be required for this clone to lyse the mAb-converted target cells. By demonstrating that a variety of different cell types can be thus converted into target cells for CTL, the approach described in this study may provide opportunities to analyze further the mechanisms by which CTL destroy target cells.     

羊轮状病毒LLR VP4单克隆抗体的制备与特性研究

目的建立抗羊轮状病毒LLR(G10P〔12])VP4特异的单克隆抗体。方法用纯化羊轮状病毒LLR免疫Balb/c小鼠,取其脾细胞与SP2/0骨髓瘤细胞进行融合,利用间接ELISA筛选阳性杂交瘤细胞,有限稀释法单克隆化,Western-blot鉴定单克隆抗体识别的抗原。结果获得4株稳定分泌抗VP4单克隆抗体的杂交瘤细胞株1B1、1B8、1F11和1G10,其中1-B1,1-B8,1-G10分泌的抗体为Ig M型,1-F11为IgG1亚类;腹水抗体效价均在1×104以上;单克隆抗体均特异性识别羊轮状病毒LLR VP4。结论1B1、1B8、1F11和1G10分泌的抗体为抗羊轮状病毒LLR VP4特异的单克隆抗体。     

Specific recognition of the product of a transferred major histocompatibility complex gene by cytotoxic T lymphocytes.

Mouse L cells transfected with a genomic clone containing the H-2Ld gene (8-5 cells) were shown to function as targets for H-2Ld-specific cytotoxic T lymphocytes (CTL). The CTL-mediated lysis of 8-5 cells was shown to be H-2Ld specific by the use of (i) CTL with restricted reactivity, (ii) unlabeled target inhibiton, and (iii) monoclonal antibody inhibiton. We also demonstrated that 8-5 cells could function as targets for antibody-plus-complement-mediated cell lysis. Specificity was confirmed by using H-2Ld-specific monoclonal antibodies. These experiments demonstrate that the gene products of a major histocompatibility complex genomic clone can be functionally expressed in a foreign cell and can mediate immunologically specific cellular interactions.     

Alloreactive cytolytic T-cell clones preferentially recognize conformational determinants on histocompatibility antigens: analysis with genetically engineered hybrid antigens.

Hybrid genes were constructed for the localization of allodeterminants on murine class I antigens recognized by antibodies and cytolytic T lymphocytes. By using deletion subclones of the H-2Kd and H-2Kk genes, homologous regions were exchanged between the two alleles. The altered genes were introduced and expressed in mouse fibroblast and fibrosarcoma cells. Cells expressing hybrid antigens were analyzed with 29 monoclonal anti-H-2Kd and anti-H-2Kk antibodies and with 150 short-term alloreactive cytolytic T-cell clones. When only the first or only the second amino-terminal domain was exchanged, most T cells and 60% of the antibodies lost their reactivity to the H-2K antigen. No T-cell clone was directed against the third extracellular domain, whereas three antibodies could bind to this domain. This implies that nearly all determinants essential for a cytolytic T-cell response or for antibody binding lie on the two external domains and are conformational structures generated by the interaction of these two domains.     

Distribution and mobility of murine histocompatibility H-2Kk antigen in the cytoplasmic membrane.

The topographical distributions and mobilities of the murine histocompatibility antigen H-2Kk and of concanavalin A (Con A) binding sites have been studied on a murine lymphoma cell line. The spatial distribution of H-2Kk antigens, the average distance between H-2Kk antigens and Con A binding sites, and the separation of different determinants on the H-2Kk antigen itself were determined by using fluorescence resonance energy-transfer measurements with a dual-laser flow sorter. From the lack of energy transfer between bound monoclonal anti-H-2Kk antibodies conjugated with fluorescein (donor) and rhodamine (acceptor), we conclude that the H-2Kk antigen exists without appreciable clustering on the cell surface. Substantial energy transfer between appropriately labeled Con A and antibodies bound to the H-2Kk antigen shows that the two populations are interspersed. Donor/acceptor pairs of monoclonal antibodies binding to different determinants on the same H-2Kk antigen exhibited a degree of energy transfer indicative of a mean separation of 8.6 nm between the sites. Time-resolved phosphorescence anisotropy measurements with anti-H-2Kk antibodies labeled with eosin or erythrosin yielded rotational mobility information for the antigen-antibody complexes on the cell membrane. The rotational correlation time of 10-20 mus and the finite residual anisotropy are compatible with an uniaxial mode of rotation of monomeric antigen around its transmembrane portion and, thus, provide additional evidence for an unclustered distribution. Capping by rabbit anti-mouse IgG immobilized the antigen-antibody complex. Fluorescence recovery after photobleaching was used to calculate an apparent lateral diffusion coefficient of 5 +/- 3 X 10(-10) cm2 . s-1 for the H-2Kk antigen labeled with fluoresceinated IgG or its corresponding Fab fragment.     

Monoclonal antibodies to novel myeloid antigens reveal human neutrophil heterogeneity.

Three cytotoxic murine monoclonal antibodies that recognize myeloid-specific antigens have been produced by immunization with normal human neutrophils or myeloblasts from a patient with acute myelomonocytic leukemia. Two of these, PMN 6 and PMN 29, are specific for neutrophils; the third monoclonal antibody, AML-2-23, is reactive with the majority of normal monocytes as well as a subpopulation of mature neutrophils. Although neutrophils from all individuals tested expressed these antigens, cytofluorographic analysis revealed that the percentage of cells bearing the PMN 6 and AML-2-23 antigens varied among individuals. Significant additional heterogeneity in the density of each antigen among antigen-bearing cells was also observed. All three antibodies efficiently mediated complement-dependent cytotoxicity of acute myelocytic leukemia cells yet were unreactive with lymphocytic leukemia cells. Neutrophil cytotoxicity was mediated by PMN 6 and PMN 29 but not by AML-2-23. On the other hand, AML-2-23, but not PMN 6 or PMN 29, was cytotoxic for normal monocytes and macrophages. These monoclonal antibodies may be of value in the study of normal neutrophil function and differentiation and may have clinical utility in diagnosis and therapy of myeloid leukemia.     

Naturally occurring cytotoxic human antibodies recognize H-2-controlled murine lymphocyte antigens.

Human sera contain cytotoxic naturally occurring (CyNa) antibodies which discriminate between lymph node cells from mice differing only at the H-2 complex. Sera from three healthy subjects (normal human sera, NH sera) and one serum from a patient with multiple sclerosis reacted with cells expressing Db, Kd, Kk, and Kp molecules, respectively. However, the following observations suggested that the binding specificity of these CyNa antibodies is to antigens that are distinct from the classical H-2 antigens: (i) the NH sera did not contain cytotoxic anti-HLA antibodies, (ii) redistribution (capping) of H-2 antigens did not induce resistance to lysis for CyNa antibodies, and (iii) individual variation was demonstrated in the expression of the murine lymphocyte antigens detected by the human CyNa antibodies. The reason for this variation appeared to be different for individual NH serum. A maternal effect influenced the expression of the murine lymphocyte antigen detected by one NH serum (anti-H-2b). The differences detected by another NH serum (anti-H-2p) appeared to be inherited, as shown by progeny testing. We hypothesize that the human CyNa antibodies may be directed against antigens controlled or modified by murine viruses (milk borne or endogenous), whose expression is under the influence of the H-2 complex, and that their production might have been stimulated by the products of human genes homologous to murine viruses.     

Fine Characterization of a Series of New Monoclonal Antibodies Directed against Glycophorin A

Objectives: Glycophorins A (GPA) and B (GPB) are the major sialoglycoproteins of the human erythrocyte (RBC) membrane. To prepare tools for the analysis of GPA and GPB, we produced a series of new monoclonal antibodies (mAbs) that identified epitopes of GPA. Methods: Seven murine monoclonal antibodies directed to glycophorin A (GPA) were fully characterized by agglutination of untreated and enzyme-treated human erythrocytes, inhibition of agglutination using chemically modified glycophorins and peptides from GPA, immunoblotting, and binding to synthetic peptides on plastic pins. Results: The antibodies identify epitopes located on four different portions of GPA: (1) NaM13-6D2 binds to the N-terminal portion of GPA and GPB carrying the N blood group antigen; (2) NaM26-3F4 recognizes the homologous portion of GPA and GPB corresponding to their amino acids 6-26; (3) NaM10-2H12, NaM16-IB10 and NaM10-6G4 are specific for the amino acid sequence 38-45 of GPA; and (4) NaM37-5F4 and NaM13-4E4 bind to the amino acid residues 119-124 located on the intracellular ponion of GPA. Conclusion: These antibodies represent precise tools to investigate GPA and related molecules in different cells and tissues.     

H-2Kk and vesicular stomatitis virus G proteins are not extensively associated in reconstituted membranes recognized by T cells.

It is shown that liposomes containing (i) a fluorescein-labeled murine histocompatibility antigen (FITC-H-2Kk) and the G protein of vesicular stomatitis virus or (ii) H-2Kk and fluorescein-labeled viral protein (FITC-G) can elicit H-2-restricted syngeneic antiviral cytotoxic T cells as assayed by 51Cr release from appropriate virus-infected target cells. Fluorescence recovery after photobleaching was used to measure the diffusion coefficients of these reconstituted proteins in four different samples: (i) FITC-H-2Kk; (ii) FITC-H-2Kk and G; (iii) FITC-G; and (iv) FITC-G and H-2Kk. The same rate of lateral diffusion (D = 1 x 10(-8) cm2/sec at 37 degrees C in 25% cholesterol/75% dimyristoylphosphatidylcholine) was obtained in every case. Both proteins, fluorescent as well as nonfluorescent, could be patched by using specific antibodies. When G was patched with antibody, FITC-H-2Kk did not copatch. When H-2Kk was patched with antibody FITC-G did not copatch. These diffusion and patching measurements rule out the possibility that these proteins have either extensive oligomeric associations or strong specific pairwise associations.     

Isolation of a T-cell clone that reacts with both antigen and anti-idiotype: evidence for anti-idiotype as internal image for antigen at the T-cell level.

T-cell lines were derived from ferredoxin nonresponder B10.D2 mice that share an idiotype expressed by a monoclonal antibody (Fd-B2) with specificity for one of the two major antigenic determinants (the C determinant) of the antigen. The T-cell line and T-cell clones derived from it release interleukin 2 not only in the presence of anti-Fd-B2 idiotype antibody but in the presence of ferredoxin. The line was shown to be major histocompatibility complex-restricted in that it would respond to the anti-idiotype and antigen only in the context of presentation by cells of the H-2d haplotype. This observation also establishes that the nonresponder status of H-2d animals cannot be attributed to a lesion at the level of antigen presentation. Analysis of the fine specificity of one idiotypic clone showed that it responded only to the anti-idiotype or products of the antigen containing the C determinant, since enzymatically degraded peptides devoid of this determinant did not stimulate these cells. Furthermore, it was found that presentation of both the antigen and the anti-idiotype to the specific clone could be blocked by the Fd-B2 monoclonal antibody.     

抗恶性疟原虫有性期单克隆抗体的制备和鉴定

以培养的恶性疟原虫ＮＦ５４（３Ｄ７）株配子体蛋白抽提液及我国云南现场采集的恶性疟原虫细胞骨架分别免疫ＢＡＬＢ／ｃ小鼠。取免疫小鼠脾细胞与ＳＰ２／０骨髓瘤细胞融合，以ＩＦＡ法筛选出８株抗恶性疟原虫有性期ＭｃＡｂ杂交瘤细胞株。经免疫球蛋白类别鉴定，６株为ＩｇＧ１（Ｍ２Ａ１０Ｃ９、Ｍ２Ｃ１Ｂ８、Ｍ４Ｃ７Ｂ１０、Ｍ４Ｇ１２Ｃ１、Ｍ５Ｂ７Ｅ６和Ｍ６Ｅ１Ｇ１１），２株为ＩｇＭ（Ｍ４Ｄ７Ｆ７和Ｍ６Ｆ４Ｄ６）。其中３株ＭｃＡｂｓ（Ｍ４Ｃ７Ｂ１０、Ｍ４Ｄ７Ｆ７和Ｍ６Ｅ１Ｇ１１）的靶抗原定位于配子体以及大滋养体和裂殖体期无性体原虫；其余５株仅定位于配子体。经Ｗｅｓｔｅｒｎ印迹试验，ＭｃＡｂ所识别的蛋白区带各异（１６－１２０ｋＤ），与已发现的有性期特异性抗原相比较，３２ｋＤ抗原国内外尚未报道。各株ＭｃＡｂ与猴疟（Ｐ．ｃｙｎｏｍｏｌｇｉ）红内期、鸡疟（Ｐ．ｇａｌｉｎａｃｅｕｍ）子孢子和杜氏利什曼原虫前鞭毛体均无交叉反应。