Absence Of Specific Antibodies Research Articles

MRNA-LNP vaccine-induced CD8+ T cells protect mice from lethal SARS-CoV-2 infection in the absence of specific antibodies

The role of CD8+ T-cells in SARS-CoV-2 pathogenesis or mRNA-LNP vaccine-induced protection from lethal COVID-19 is unclear. Using mouse-adapted SARS-CoV-2 virus (MA30) in C57BL/6 mice, we show that CD8+ T-cells are unnecessary for the intrinsic resistance of female or the susceptibility of male mice to lethal SARS-CoV-2 infection. Also, mice immunized with a di-proline prefusion-stabilized full-length SARS-CoV-2 Spike (S-2P) mRNA-LNP vaccine, which induces Spike-specific antibodies and CD8+ T-cells specific for the Spike-derived VNFNFNGL peptide, are protected from SARS-CoV-2 infection-induced lethality and weight loss, while mice vaccinated with mRNA-LNPs encoding only VNFNFNGL are protected from lethality but not weight loss. CD8+ T-cell depletion ablates protection in VNFNFNGL but not in S-2P mRNA-LNP-vaccinated mice. Therefore, mRNA-LNP vaccine-induced CD8+ T-cells are dispensable when protective antibodies are present but essential for survival in their absence. Hence, vaccine-induced CD8+ T-cells may be critical to protect against SARS-CoV-2 variants that mutate epitopes targeted by protective antibodies.

Imaging flow cytometry of tumoroids: A new method for studying GPCR expression.

Fluorescence confocal microscopy is commonly used to analyze the regulation membrane proteins expression such as G protein-coupled receptors (GPCRs). With this approach, the internal movement of GPCRs within the cell can be observed with a high degree of resolution. However, these microscopy techniques led to complex and time-consuming analysis and did not allow a large population of events to be sampled. A recent approach termed imaging flow cytometry (IFC), which combines flow cytometry and fluorescence microscopy, had two main advantages to study the regulation of GPCRs expression such as orexins receptors (OXRs): the ability (1) to analyze large numbers of cells and; (2) to visualize cell integrity and fluorescent markers localization. Here, we compare these two technologies using the orexin A (OxA) ligand coupled to rhodamine (OxA-rho) to investigate anti-tumoral OX1R expression in human digestive cancers. IFC has been adapted for cancer epithelial adherent cells and also to 3D cell culture tumoroids which partially mimic tumoral structures. In the absence of specific antibody, expression of OX1R is examined in the presence of OxA-rho. 2D-culture of colon cancer cells HT-29 exhibits a maximum level of OX1R internalization induced by OxA with 19% ± 3% colocalizing to early endosomes. In 3D-culture of HT-29 cells, internalization of OX1R/OxA-rho reached its maximum at 60 min, with 30.7% ± 6.4% of OX1R colocalizing with early endosomes. This is the first application of IFC to the analysis of the expression of a native GPCR, OX1R, in both 2D and 3D cultures of adherent cancer cells.

Serological and molecular evidence of chikungunya virus infection among febrile outpatients seeking healthcare in Northern Malawi

ABSTRACT Introduction: Despite global evidence of chikungunya fever (CHIKF) in humans that is caused by chikungunya virus (CHIKV), little is known about the occurrence of CHIKF in Malawi. This study was conducted to determine the seroprevalence of CHIKF and to molecularly confirm the presence of CHIKV ribonucleic acid (RNA) among febrile outpatients seeking health care at Mzuzu Central Hospital in the Northern Region of Malawi. Methods: Enzyme-immunosorbent assay (ELISA) was used to detect the presence or absence of specific antibodies against CHIKV. Reverse transcription-polymerase chain reaction (RT-PCR) was conducted on randomly selected anti-CHIKV IgM-positive samples to detect CHIKV RNA. Results: Out of 119 CHIKF suspected samples analyzed, 73 tested positive for anti-CHIKV IgM antibodies, with an overall seroprevalence of 61.3%. Most of the CHIKV infected individuals presented with joint pain, abdominal pain, vomiting and nose bleeding with seroprevalence of 45.2%, 41.1%, 16.4% and 12.3%, respectively. All the randomly selected samples that were positive for CHIKV anti-IgM by ELISA had detectable CHIKV RNA by RT-PCR. Conclusion: The presence of anti-CHIKV IgM antibodies suggests the presence of recent CHIKV infection. We therefore recommend for the inclusion of CHIKF as the differential diagnosis in febrile ill patients in Mzuzu city, Malawi.

D-Alanylation of Lipoteichoic Acids in Streptococcus suis Reduces Association With Leukocytes in Porcine Blood.

Streptococcus suis (S. suis) is a common swine pathogen but also poses a threat to human health in causing meningitis and severe cases of streptococcal toxic shock-like syndrome (STSLS). Therefore, it is crucial to understand how S. suis interacts with the host immune system during bacteremia. As S. suis has the ability to introduce d-alanine into its lipoteichoic acids (LTAs), we investigated the working hypothesis that cell wall modification by LTA d-alanylation influences the interaction of S. suis with porcine blood immune cells. We created an isogenic mutant of S. suis strain 10 by in-frame deletion of the d-alanine d-alanyl carrier ligase (DltA). d-alanylation of LTAs was associated with reduced phagocytosis of S. suis by porcine granulocytes, reduced deposition of complement factor C3 on the bacterial surface, increased hydrophobicity of streptococci, and increased resistance to cationic antimicrobial peptides (CAMPs). At the same time, survival of S. suis was not significantly increased by LTA d-alanylation in whole blood of conventional piglets with specific IgG. However, we found a distinct cytokine pattern as IL-1β but not tumor necrosis factor (TNF)-α levels were significantly reduced in blood infected with the ΔdltA mutant. In contrast to TNF-α, activation and secretion of IL-1β are inflammasome-dependent, suggesting a possible influence of LTA d-alanylation on inflammasome regulation. Especially in the absence of specific antibodies, the association of S. suis with porcine monocytes was reduced by d-alanylation of its LTAs. This dltA-dependent phenotype was also observed with a non-encapsulated dltA double mutant indicating that it is independent of capsular polysaccharides. High antibody levels caused high levels of S. suis—monocyte—association followed by inflammatory cell death and strong production of both IL-1β and TNF-α, while the influence of LTA d-alanylation of the streptococci became less visible. In summary, the results of this study expand previous findings on d-alanylation of LTAs in S. suis and suggest that this pathogen specifically modulates association with blood leukocytes through this modification of its surface.

The Anti-SARS-CoV-2 Antibody Response in a Centenarian Woman: A Case of Long-Term Memory?

SARS-CoV-2 is the virus responsible for the COVID-19 pandemic, causing respiratory syndrome and other manifestations. The clinical consequences of the SARS-CoV-2 infection are highly heterogeneous, ranging from asymptomatic and mild to severe and fatal conditions, with the highest mortality rate reached among elderly people. Such heterogeneity appears strongly influenced by the host immune response, which in turn is profoundly affected by aging. In fact, the occurrence of a low-grade inflammation and a decline in specific immune defense is generally reported in older people. Although the low ability of B cells to provide primary and secondary specific responses with a consequent increase in susceptibility to and severity of virus infections is generally described in elderly people, we would like to present here the particular case of a 100-year-old woman, who recovered well from COVID-19 and developed a long-term memory against SARS-CoV-2. Following the infection, the patient’s blood was tested with both a classical ELISA and a specific Cell-ELISA addressed to measure the anti-spike S1 specific IgG released in plasma or produced in vitro by memory B cells, respectively. While showing negative on classical serological testing, the patient’s blood was positive in Cell-ELISA up to 1 year after the infection. Our observation highlights a potential mechanism of B cell-dependent, long-term protection in response to SARS-CoV-2 infection, suggesting that in a case of successful aging, the absence of specific antibodies in serum does not necessarily mean the absence of immune memory.

Mouse lung tissue resident memory CD8 T cells exhibit unique metabolic profiles

Abstract Tissue resident memory T cells (TRM) confer protection against a variety of illnesses ranging from infections to cancers. As demonstrated in the skin, transfer of TRM from immune to naïve hosts is sufficient to prevent herpes simplex virus type 1 (HSV) associated pathology. Comparably, the presence of antigen-specific TRM in a mouse model of melanoma can limit tumor growth. In the lungs, TRM offer critical for protection against influenza, even in the absence of specific antibodies. In the skin, the functions of different CD8 T cell subsets have been linked with distinct metabolic pathways. Skin TRM display a unique metabolic profile to other CD8 memory cells subsets. They undergo enhanced fatty acid oxidation and oxidative phosphorylation as compared with circulating memory cells and naïve cells. However, it is unclear if this programming is universal between all TRM subsets and holds true in the lung. We hypothesize that lung TRM with different integrin profiles will utilize unique metabolic programs. Metabolic profiles of lung CD8 T cells were assessed using a variety of functional metabolic assays including glucose analogue uptake, fatty acid uptake, and oxidative phosphorylation as assessed by fluorescent dyes measured by flow cytometry. Nonfunctional assays including neutral lipid storage, mitochondrial mass, and mRNA transcript levels of key metabolic genes were also assessed via flow cytometry using fluorescent dyes, while transcripts were quantified via reverse transcription quantitative polymerase chain reaction (RT-qPCR). Our findings indicate that lung TRM show enhanced fatty acid oxidation, more neutral lipid storage, and less glucose uptake than their non-TRM T cells in our influenza mouse model.

Not just antibodies: B cells and T cells mediate immunity to COVID-19.

Recent reports that antibodies to SARS-CoV-2 are not maintained in the serum following recovery from the virus have caused alarm. However, the absence of specific antibodies in the serum does not necessarily mean an absence of immune memory. Here, we discuss our current understanding of the relative contribution of B cells and T cells to immunity to SARS-CoV-2 and the implications for the development of effective treatments and vaccines for COVID-19. Here, Cox and Brokstad briefly discuss T cell- and B cell-mediated immunity to SARS-CoV-2, stressing that a lack of serum antibodies does not necessarily equate with a lack of immunity to the virus.

Polymyalgische Beschwerden: Steckt ein Tumor dahinter?

A 61-year-old caucasian male presenting with pain in arms and thighs, weight loss of 10 kg in the last 3 months, and subfebrile temperatures, also had abdominal pain. In addition, he reported that his abdominal circumference had increased over the last two years. Based on patient's complaints and the presence of elevated inflammatory markers in the absence of specific antibodies or peripheral synovitis a clinical diagnosis of polymyalgia rheumatica was made. The patient had an immediate clinical response to 20 mg prednisone/day, and inflammatory markers normalized. An abdominal MRI showed no signs of polymyalgia rheumatica but revealed a lipomatous mass with displacement of the adjacent intraabdominal and intrapelvine organs. Histologically a highly differentiated retroperitoneal liposarcoma was described. The patient underwent a complete resection of the tumor. Because the liposarcoma was highly differentiated, the oncologists were reluctant to use any additional therapy. The diagnostic work up of polymyalgic complaints should also include rare paraneoplastic diseases.

Novel biosensing system for the simultaneous multiplex fluorescent determination of catecholamines and their metabolites in biological liquids

A novel original biosensing system for the simultaneous multiplex determination of general markers of catecholamine-producing diseases – catecholamines (dopamine, epinephrine, norepinephrine) and their metabolites (homovanillic and vanillylmandelic acids) in biological liquids without preliminary separation of analytes, in the absence of specific antibodies and receptors and with minimum pretreatment of a samples has been developed. This outstanding approach includes the unique combination of obtaining highly fluorescent derivatives of the analytes as a result of their interaction with two different amines ̶ benzylamine and 1,2-diphenylethylenediamine in the presence of peroxidase as a catalyst, with the application of first-order derivative fluorescence spectroscopy for the resolution of their spectra. Fluorescence is measured in 96-well microplates, which wells contain a bio-recognizing film consisted of horseradish peroxidase immobilized in the polymer chitosan. Spectra of the solutions are recorded in the range 400–500 nm (λex ∼ 305–356 nm). The proposed procedures provide sensitive (in the range of 3–200 nM), selective, and reproducible (RSDs ≤ 1%, n = 6) multiplex determination of the catecholamines and their metabolites in biological liquids were successfully applied for the rapid simultaneous (20 samples per 15–30 min) screening of human urine and mice blood plasma. The validated results showed good linearity, precision, accuracy and selectivity of this method.

Functional Analysis Reveals Geographical Variation in Inhibitory Immune Responses Against a Polymorphic Malaria Antigen.

Plasmodium falciparum reticulocyte-binding protein homologue 2b (PfRh2b) is an invasion ligand that is a potential blood-stage vaccine candidate antigen; however, a naturally occurring deletion within an immunogenic domain is present at high frequencies in Africa and has been associated with alternative invasion pathway usage. Standardized tools that provide antigenic specificity in in vitro assays are needed to functionally assess the neutralizing potential of humoral responses against malaria vaccine candidate antigens. Transgenic parasite lines were generated to express the PfRh2b deletion. Total immunoglobulin G (IgG) from individuals residing in malaria-endemic regions in Tanzania, Senegal, and Mali were used in growth inhibition assays with transgenic parasite lines. While the PfRh2b deletion transgenic line showed no change in invasion pathway utilization compared to the wild-type in the absence of specific antibodies, it outgrew wild-type controls in competitive growth experiments. Inhibition differences with total IgG were observed in the different endemic sites, ranging from allele-specific inhibition to allele-independent inhibitory immune responses. The PfRh2b deletion may allow the parasite to escape neutralizing antibody responses in some regions. This difference in geographical inhibition was revealed using transgenic methodologies, which provide valuable tools for functionally assessing neutralizing antibodies against vaccine-candidate antigens in regions with varying malaria endemicity.

The complement receptor CRIg dominates clearance of intravascular parasites during African trypanosome infection

Abstract Kupffer cells (KCs), the liver resident macrophages, play a prominent role in clearance of blood-borne pathogens. The mechanisms by which KCs engulf circulating parasites such as African trypanosomes have yet to be determined. Here we show that depletion of KCs resulted in uncontrolled parasitemia in mice infected with Trypanosoma congolense, indicating the essential role of KCs in clearance of intravascular parasites. By using intravital microscopy, we further investigated the dynamics of interactions of KCs with circulating T. congolense in real-time. The results demonstrate that KCs catch circulating parasites in the presence of specific IgM and IgG antibodies, but fail to do so in the absence of specific antibodies. Complement C3 is critically involved in the capture of the parasites mediated by both IgM and IgG. We further demonstrate that complement receptor CRIg, but not CR3, is crucial for KCs to catch the parasites. In support of the findings gained by in vivo imaging, mice deficient of C3 or CRIg, but not CR3, could not control the first wave of parasitemia, and had a dramatic reduced survival time compared to wild-type mice following infection with T. congolense (10 days vs over 100 days). Collectively, our data demonstrate that CRIg play a dominant role in the clearance of circulating parasites by KCs in an antibody dependent manner.

Stimulation of Phagocytosis and Production of Antibodies against Canine Herpesvirus Type 1 by Pidotimod (Adimod<sup>TM</sup>)

Neutrophils are the most important circulating phagocytes. Circulating mono-cytes and precursors of tissue macrophages also have the ability to phagocytize. Pidotimod (ADIMODTM) exerts immunostimulatory and immunoregulatory effects through the stimulation and regulation of cellular immune responses by lymphocytes Canine herpesvirus (CHV) mainly affect puppies between the first and second weeks of age, causing high morbidity in the litter. To date, there is only one commercial vaccine in Europe to prevent disease. In this work, inactivated CHV cultures were inoculated in rabbits, adsorbed and not adsorbed to chitosan nanoparticles. Phagocytosis in the presence or absence of specific antibodies was measured. Response of virus neutralizing antibodies was also evaluated. AdimodTM enhanced the nonspecific and specific phagocytotic response. The association of the virus to the nanoparticles increased the phagocytic ability of blood cells; however, AdimodTM alone had a greater effect on phagocytic activity and generated a stronger immune response that corresponded to the increased phagocytosis (p TM was used.

Proximity Ligation Assay to Quantify Foxp3 Acetylation in Regulatory T Cells.

Determining protein acetylation by immunoprecipitation and immunoblotting can be challenging, especially if the tissue of interest is low in quantity, and when good quality acetylation site-specific antibodies are not available. Proximity ligation assays allow a sensitive and quantitative method to assess Foxp3 acetylation in regulatory T cells, with as little as 1.5 × 105 cells within two days turnaround time. This method is of potential use in other similar scenarios, when post-translational modifications of a protein of interest need to be determined with only a small amount of sample and in the absence of specific antibodies that can assess the post-translational modification in the protein of interest.

Detection of Phospho-KRAS by Electrophoretic Mobility Change in Human Cell Lines and in Tumor Samples from Nude Mice Grafts

[Abstract] KRAS is the oncogene most frequently mutated in human solid tumors especially in pancreas, colon, small intestine, biliary tract and lung. We have recently demonstrated that oncogenic KRAS needs S181 phosphorylation to fully display its oncogenic features suggesting its inhibition as a therapeutic treatment against KRAS-driven tumors. Due to the importance to detect KRAS phosphorylation in human tumors and the absence of specific antibodies against phosphorylated KRAS, we developed a new protocol based on the Phos-tag SDS methodology to detect this post-translational modification for KRAS. Phos-tag is a molecule that binds specifically to phosphorylated proteins, decreasing their migration speed in SDS-PAGE and allowing its separation from the non-phosphorylated forms.

CD4+ T cells limit the damage in influenza

Why do some influenza infections cause fatal disease and others barely a sniffle? Although viral virulence can vary, the immunological history of the host is also important. A new study in humans suggests that CD4+ T lymphocytes activated during previous infections can limit disease severity in the absence of specific antibodies (pages 274–280).

Serological and Virological Analysis of Maternal and Fetal Blood Samples in Prenatal Human Parvovirus B19 Infection

Intrauterine parvovirus B19 (B19V) infection can be asymptomatic or may cause severe fetal complications. Information on serological and virological findings of infection in the fetus is scarce. We determined B19V-DNA and anti-B19V antibodies in maternal and fetal blood samples obtained from 41 pregnancies that were complicated by prenatal B19V infection. Most fetuses presented with moderate to severe anemia or hydrops. At the time of fetal blood sampling, all mothers were B19V-DNA positive and B19V-IgG positive. B19V-IgM was detected in 95% of maternal blood samples. B19V-DNA, B19V-IgM, and B19V-IgG were detected in 100%, 28%, and 24% of fetal blood samples, respectively. The probability of a positive B19V-IgG or B19V-IgM finding in fetal blood increased with gestational age. B19V-IgG levels in maternal blood did not correlate with the likelihood of a positive B19V-IgG test in the fetus. The presence of B19V-IgG in fetal blood was accompanied by lower B19V-DNA levels and less severe clinical findings. The lack of B19V-IgG in fetuses with B19V-derived anemia or hydrops is most likely due to a limited materno-fetal transfer of IgG and a poor fetal antibody response. Fetal B19V infection is poorly controlled in the absence of specific antibodies.

Adherence of Staphylococcus epidermidis to human endothelial cells is associated with a polysaccharidic component of its extracellular mucous layer

Bacterial adherence to eukaryotic cells is highly contributing to microbial pathogenesis. Bacterial adhesins, macromolecules, and glycosaminoglycan chains of the endothelial cell surface have been implicated in staphylococcal attachment. Our research group has isolated an antigenic polysaccharidic component of Staphylococcus epidermidis extracellular layer, known as 20-kDa PS (PS), and showed that antibodies against this polysaccharide protect from infections. Therefore, the role of PS in S. epidermidis adherence to endothelial cells was studied. For this purpose we examined the impact of PS on the ability of two S. epidermidis strains (a PS-producing and a non-PS-producing strain) to adhere to human endothelial cells in the presence or absence of specific antibodies to this polysaccharide. Hence, it is established that exogenous chondroitin sulfate (CS) decreases, in part, the S. epidermidis' attachment to endothelial cells and the antagonistic binding effect of CS and PS was also studied. The results obtained demonstrate that PS facilitates the adherence of S. epidermidis to both strains. CS abolished the PS-induced adherence in PS‐producing strain and partially in the non-PS-producing one. Conclusively, the adherence of S. epidermidis to human endothelial cells is associated with its extracellular PS component and it is suggested that the bacterial binding via glycosaminoglycan chains is an important mechanism underlining the PS-induced binding to endothelial cells.

Effects of integrins on laminin chemotaxis by hepatocellular carcinoma cells

To quantitatively evaluate the effects of integrins alpha1beta1, alpha2beta1, alpha3beta1, alpha4beta1, alpha5beta1, and alpha6beta1 on the chemotaxis of hepatocelluar carcinoma (HCC) cell line SMMC-7721 to laminin (LN). A modified dual-micropipette system was used to dynamically and quantitatively monitor the formation of pseudopod protrusion of HCC cells toward LN in the presence or absence of specific antibodies against integrins alpha1, alpha2, alpha3, alpha4, alpha5, alpha6, and beta1. Additionally, the expression levels of different integrin subunits on the surface of the cells were determined via flow cytometry analysis. In response to equal concentrations of LN in both micropipettes, HCC cells form symmetrical pseudopod protrusions on both sides. Addition of antibodies against alpha3, alpha6, or beta1 into one micropipette leads to significant reduction of pseudopod formation on that side, while antibodies against alpha1, alpha2, alpha4, and alpha5 do not affect the symmetrical formation of pseudopods in either micropipette. The percentages of HCC cells positive for expression of integrins alpha1, alpha2, alpha3, alpha4, alpha5, alpha6, and beta1 were 95.07, 23.17, 95.55, 2.47, 34, 14.29, and 95.78%, respectively. Integrins alpha3beta1 and alpha6beta1 are important cell surface receptors that mediate the chemotaxis of HCC cells toward LN.

Chapter 5: Strategies to prevent transmission of porcine endogenous retroviruses

Xenotransplantation using porcine cells, tissues, or organs may offer a potential solution for the shortage of allogeneic human organs. Prior to the clinical use of porcine xenotransplants, three main hurdles must be overcome: immunologic rejection, physiologic incompatibility, and risk of transmission of porcine pathogens. Designated pathogen-free breeding of pigs can prevent transmission of most porcine microbes. However, this is not possible in the case of porcine endogenous retroviruses (PERV), which are integrated in the pig genome and can infect human cells in vitro. In order to assess the probability of transmission of PERV, a careful screening of the source pig herd is recommended. Two types of PERV are present in pigs, human-tropic PERV-A and PERV-B, which are both present in the genome of all pigs, and PERV-C, which is not ubiquitous and infects only pig cells. In addition to these viruses, recombinant PERV-A/C viruses have recently been described that (i) are able to infect human cells; (ii) are characterized by high titre replication; and (iii) are associated with proviruses that are de novo integrated in the DNA of somatic pig cells, but not yet in the pig germ line. The risks presented by PERV-A/C recombinant viruses could easily be eliminated by using pigs not containing PERV-C in their germ line, thereby effectively preventing recombination with PERV-A. Selection of PERV-C-free animals, if possible, therefore reduces the risk of PERV-A/C transmission to humans. Although it is unclear whether PERV-C may be transmitted in vivo from pig-to-pig, an infection of PERV-C-free animals with this virus may be prevented. To select pigs with low-level expression of PERV-A and PERV-B, it is recommended to apply assays based on real-time reverse transcriptase polymerase chain reaction (RT-PCR), which enables discrimination between pigs with high-level expression and low-level expression. Screening xenotransplant recipients for PERV transmission can be done in a number of ways. Provirus integration and PERV expression could theoretically be detected in peripheral blood mononuclear cells using PCR and RT-PCR. However, as the cells in which PERV replicates are still unknown, it is unclear whether this will be a reliable approach. Applying sufficiently sensitive assays to differentiate between transmission and chimerism is recommended. As can be commonly observed after retrovirus infection, the detection of virus-specific antibodies may indicate infection; however, the possibility of abortive infection, antigen exposure without infection, or cross-reactive response must be considered and explored as alternative explanations. On the other hand, it remains unclear whether the absence of specific antibodies indicates the absence of such infection, in particular if recipients of a xenotransplantation product are under chronic immunosuppression that could prevent antibody formation. Antibodies may be detected by Western blot or ELISA, using purified virus or recombinant viral proteins as antigens. Finally, it may be possible to detect cross-species PERV transmission by evaluating cells from the recipient for their in vitro potential of transmission to specified target cells (the human renal epithelial 293 cell line being the best example). There is no in vivo animal model for cross-species PERV transmission, and therefore it is not possible to validate monitoring assays for PERV transmission in an in vivo situation. Finally, virus safety of xenotransplantation is a fast-developing field, and new experimental findings will change existing strategies and introduce new ones.

Immunoparasitological evaluation of Trichinella spiralis infection during human pregnancy: a small case series

The clinical background, vertical transmission of Trichinella spiralis and helminthotoxic activity of sera were evaluated in four pregnant women infected at different gestation times. Parasitological and immunological parameters were analyzed. Patients presented clinical symptoms and a seroconversion pattern typical of Trichinella infection, independently of the gestation trimester. All patients, including one patient treated with mebendazole, gave birth to healthy infants at full term. Studies performed in placentas and umbilical cords by artificial digestion and/or immunofluorescence did not reveal the presence of parasites or parasite antigens. Specific antibodies were found in placental extraction and maternal sera by immunofluorescence, ELISA and/or immunoelectrotransfer blot assay. Specific IgG, IgE and IgA were found in the umbilical cord sera. One umbilical cord demonstrated immune-complexed specific IgM and circulating parasite antigens were found in the corresponding infant up to 10 months after birth. Patients' sera were able to kill newborn larvae in cytotoxicity assays, even in the absence of specific antibodies; this effect was abrogated by mifepristone. The results suggest that in human trichinellosis during pregnancy there is an enhanced helminthotoxicity to newborn larvae, dependent in part on progesterone, leading to a mild or moderate course of the infection. The results also indicate that the transplacental passage of migrating larvae is possible.