scholarly journals Serological Expression Cloning of Novel Immunoreactive Antigens of Babesia microti

2000 ◽  
Vol 68 (5) ◽  
pp. 2783-2790 ◽  
Author(s):  
Michael J. Lodes ◽  
Raymond L. Houghton ◽  
Elizabeth S. Bruinsma ◽  
Raodoh Mohamath ◽  
Lisa D. Reynolds ◽  
...  

ABSTRACT Increased recognition of the prevalence of human babesiosis in the United States, together with rising concern about the potential for transmission of this infection by blood transfusion, has provided motivation to develop definitive serologic and molecular tests for the causative agent, Babesia microti. To develop more sensitive and specific assays for B. microti, we screened a genomic expression library with patient serum pools. This screening resulted in the identification of three classes of novel genes and an additional two novel, unrelated genes, which together encode a total of 17 uniqueB. microti antigens. The first class (BMN1-2 family) of genes encodes seven closely related antigens with a degenerate six-amino-acid repeat that shows limited homology toPlasmodium sp. merozoite and sporozoite surface antigens. A second class (BMN1-8 family) of genes encodes six related antigens, and the third class (BMN1-17 family) of genes encodes two related antigens. The two remaining genes code for novel and unrelated sequences. Among the three classes of antigens and remaining novel sequences, five were chosen to code for the most immunodominant antigens (BMN1-2, -9, -15, and -17 and MN-10). Western blot analysis with the resulting recombinant proteins indicated that these antigens were targets of humoral immune responses during B. microti infection in humans.

Author(s):  
Adi V Gundlapalli ◽  
Reynolds M Salerno ◽  
John T Brooks ◽  
Francisco Averhoff ◽  
Lyle Petersen ◽  
...  

Abstract Background There is a need for validated and standardized SARS-CoV-2 quantitative IgG and neutralization assays that can be used to understand the immunology and pathogenesis of SARS-CoV-2 infection and support the COVID-19 pandemic response. Methods Literature searches were conducted to identify English language publications from peer-reviewed journals and pre-prints from January 2020 through October 12, 2020. Relevant publications were reviewed for mention of IgG or neutralization assays for SARS-CoV-2, or both, and the methods of reporting assay results. Results Quantitative SARS-CoV-2 IgG results have been reported from a limited number of studies; most studies used in-house laboratory-developed tests in limited settings, and only two semi-quantitative tests have received U.S. Food and Drug Administration (FDA) Emergency Use Authorization (EUA). As of October12, 2020, there are no neutralization assays with FDA EUA. Relatively few studies have attempted correlation of quantitative IgG titers with neutralization results to estimate surrogates of protection. The number of individuals tested is small compared to the magnitude of the pandemic and persons tested are not representative of disproportionately affected populations. Methods of reporting quantitative results are not standardized to enable comparisons and meta-analyses. Conclusions Lack of standardized SARS-CoV-2 quantitative IgG and neutralization assays precludes comparison of results from published studies. Inter-assay and inter-laboratory validation and standardization of assays will support efforts to better understand antibody kinetics and longevity of humoral immune responses post-illness, surrogates of immune protection, and vaccine immunogenicity and efficacy. Public-private partnerships could facilitate realization of these advances in the United States and world-wide.


Pathogens ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 1447
Author(s):  
Abhinav Kumar ◽  
Jane O'Bryan ◽  
Peter J. Krause

Babesiosis is an emerging tick-borne disease caused by intraerythrocytic protozoa that are primarily transmitted by hard-bodied (Ixodid) ticks and rarely through blood transfusion, perinatally, and organ transplantation. More than 100 Babesia species infect a wide spectrum of wild and domestic animals worldwide and six have been identified as human pathogens. Babesia microti is the predominant species that infects humans, is found throughout the world, and causes endemic disease in the United States and China. Babesia venatorum and Babesia crassa-like agent also cause endemic disease in China. Babesia divergens is the predominant species in Europe where fulminant cases have been reported sporadically. The number of B. microti infections has been increasing globally in recent decades. In the United States, more than 2000 cases are reported each year, although the actual number is thought to be much higher. In this review of the epidemiology of human babesiosis, we discuss epidemiologic tools used to monitor disease location and frequency; demographics and modes of transmission; the location of human babesiosis; the causative Babesia species in the Americas, Europe, Asia, Africa, and Australia; the primary clinical characteristics associated with each of these infections; and the increasing global health burden of this disease.


Pathogens ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 1563
Author(s):  
Scott Meredith ◽  
Miranda Oakley ◽  
Sanjai Kumar

The biology of intraerythrocytic Babesia parasites presents unique challenges for the diagnosis of human babesiosis. Antibody-based assays are highly sensitive but fail to detect early stage Babesia infections prior to seroconversion (window period) and cannot distinguish between an active infection and a previously resolved infection. On the other hand, nucleic acid-based tests (NAT) may lack the sensitivity to detect window cases when parasite burden is below detection limits and asymptomatic low-grade infections. Recent technological advances have improved the sensitivity, specificity and high throughput of NAT and the antibody-based detection of Babesia. Some of these advances include genomics approaches for the identification of novel high-copy-number targets for NAT and immunodominant antigens for superior antigen and antibody-based assays for Babesia. Future advances would also rely on next generation sequencing and CRISPR technology to improve Babesia detection. This review article will discuss the historical perspective and current status of technologies for the detection of Babesia microti, the most common Babesia species causing human babesiosis in the United States, and their implications for early diagnosis of acute babesiosis, blood safety and surveillance studies to monitor areas of expansion and emergence and spread of Babesia species and their genetic variants in the United States and globally.


Author(s):  
Suzanne Moshier ◽  
William O'Dell ◽  
Raychel Watkins ◽  
Aelita Pinter

In a review of the parasites of Microtus, Timm (1985) lists no protozoan endoparasites whatsoever for this genus. The role of parasitism, whether macro- or microparasites, and whether endo- or ectoparasites, in the demographic machinery of microtines is poorly understood. Timm (1985) astutely observes that one of the most challenging and fruitful directions of future research with Microtus will be the statistical quantification of the cost of parasitism. In addition, since humans in the Grand Teton National Park may encounter protozoon parasites that are potentially pathogenic to humans, it is useful to characterize the occurrence and biology of such organisms in the parko Babesia microti, a parasitic protozoon, is transmitted by a tick vector and reproduces in the erythrocytes of its mammalian host. Initially, Babesia was thought to be restricted to small mammals; however, in 1970 the first human cases were diagnosed in residents of Nantucket Island, Massachusetts (Western et al, 1970). Over 200 cases of human babesiosis have been documented worldwide. In the United States, human babesiosis is caused by B. microti. Most of these cases have occurred in the eastern United States. The earliest report of an organism that fits the description of Babesia in human erythrocytes is that of Wilson et al. (1904), who found an unknown organism in human erythrocytes while investigating the cause of Rocky Mountain Spotted fever. Documented cases of babesiosis in many areas of the United States are increasing (Steketee et al, 1985). As humans insert themselves into places where they have historically been present only occasionally, they often contract new diseases. A second protozoon parasite, Hepatozoon sp., which is widespread in small mammals in Europe, is also found in reptiles throughout the world. The record of Hepatozoon in North American small mammals is not extensive. Fewer than 10 species of mammals have been shown to harbor Hepatozoon parasites. Like Babesia, Hepatozoon is a two-host parasite. Unlike Babesia, for which the intermediate host is always a tick, the intermediate host in the Hepatozoon may be a tick, a mite, a flea, or a mosquito. The method of transmission by the vector also differs in the two parasites. Babesia is transmitted in saliva when the tick bites, whereas Hepatozoon infection requires the vertebrate host to swallow the vector. In our 1994 studies, we sought to extend knowledge of these two parasites. The specific objectives for 1994 were: to sample specific populations of M. molltanus, in which we have previously documented Hepatozoon infections, to determine whether there are differences in the infection rates at different study sites in the park; to search for the vector of Hepatozoon sp. infections in M. montanus by examining ectoparasites; to collect and rear ticks from M. montanus for use in R microti transmission studies; and to complete a comparison of the spleen histology of babesiosis in infected and uninfected laboratory animals, with inclusion of data from wild animals, as available. Our long-term objectives are to document the effects and cost of parasitism on vole populations and to determine the potential of small mammals of Grand Teton National Park to serve as reservoirs of human parasites.


Pathogens ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 1176
Author(s):  
Evan M. Bloch ◽  
Peter J. Krause ◽  
Laura Tonnetti

Babesia are tick-borne intra-erythrocytic parasites and the causative agents of babesiosis. Babesia, which are readily transfusion transmissible, gained recognition as a major risk to the blood supply, particularly in the United States (US), where Babesia microti is endemic. Many of those infected with Babesia remain asymptomatic and parasitemia may persist for months or even years following infection, such that seemingly healthy blood donors are unaware of their infection. By contrast, transfusion recipients are at high risk of severe babesiosis, accounting for the high morbidity and mortality (~19%) observed in transfusion-transmitted babesiosis (TTB). An increase in cases of tick-borne babesiosis and TTB prompted over a decade-long investment in blood donor surveillance, research, and assay development to quantify and contend with TTB. This culminated in the adoption of regional blood donor testing in the US. We describe the evolution of the response to TTB in the US and offer some insight into the risk of TTB in other countries. Not only has this response advanced blood safety, it has accelerated the development of novel serological and molecular assays that may be applied broadly, affording insight into the global epidemiology and immunopathogenesis of human babesiosis.


2000 ◽  
Vol 38 (1) ◽  
pp. 362-368
Author(s):  
M. J. Homer ◽  
E. S. Bruinsma ◽  
M. J. Lodes ◽  
M. H. Moro ◽  
S. Telford ◽  
...  

ABSTRACT Human babesiosis in the United States is caused predominantly by Babesia microti , a tick-transmitted blood parasite. Improved testing methods for the detection of infection with this parasite are needed, since asymptomatic B. microti infection represents a potential threat to the blood supply in areas where B. microti is endemic. We performed immunoscreening of an expression library of genomic DNA from a human isolate of B. microti (strain MN1). Among 17 unique immunoreactive clones, we identified 9 which represent a related family of genes with little sequence homology to other known sequences but with an architecture resembling that of several surface proteins of Plasmodium . Within this family, a tandem array of a degenerate six-amino-acid repeat (SEAGGP, SEAGWP, SGTGWP, SGTVGP) was found in various lengths between relatively well conserved segments at the N and C termini. In order to examine within-clone variation, we developed a PCR protocol for direct recovery of a specific bmn1-6 homologue directly from 30 human blood isolates, 4 corresponding hamster isolates, and 5 geographically corresponding Peromyscus leucopus (white-footed mouse) isolates. Isolates from the hamsters had the same sequences as those found in the corresponding human blood, suggesting that genetic variation of bmn1-6 does not occur during passage. However, clones from different patients were often substantially different from each other with regard to the number and location of the degenerate repeats within the bmn1-6 homologue. Moreover, we found that strains that were closely related geographically were also closely related at the sequence level; nine patients, all from Nantucket Island, Mass., harbored clones that were indistinguishable from each other but that were distinct from those found in other northeastern or upper midwestern strains. We conclude that considerable genetic and antigenic diversity exists among isolates of B. microti from the United States and that geographic clustering of subtypes may exist. The nature of the bmn1-6 gene family suggests a mechanism of antigenic variation in B. microti that may occur by recombination, differential expression, or a combination of both mechanisms.


Parasitology ◽  
2003 ◽  
Vol 127 (4) ◽  
pp. 301-309 ◽  
Author(s):  
H. K. GOETHERT ◽  
S. R. TELFORD

Babesia microti(Apicomplexa: Piroplasmida) has historically been considered a common parasite of Holarctic rodents. However, human babesiosis due to this species has generally been limited to the northeastern seaboard of the United States and Minnesota and Wisconsin. The absence of reports ofB. microtibabesiosis from sites where the agent is enzootic, such as in western Europe, remains unexplained. Previous work focusing on the 18S rDNA demonstrates little sequence diversity among samples from allopatric host populations across a wide geographical area. It may be that genetic diversity is underestimated due to sample size or the gene analysed. Accordingly, we collected blood or spleen samples from American or Eurasian animals with parasites that were morphologically consistent withB. microti, amplified the 18S rDNA andbeta-tubulin gene, and conducted phylogenetic analysis. Surprisingly, what was considered to be ‘B. microti’ by microscopy appears to be a diverse species complex. We identify 3 distinct clades within this complex, including parasites from non-rodent hosts. Rodent parasites comprise 2 clades, one representing zoonotic isolates, and the other apparently maintained in microtine rodents, and therefore their morphological detection within animals from a site does not necessarily imply a risk to public health.


2017 ◽  
Vol 55 (10) ◽  
pp. 2903-2912 ◽  
Author(s):  
Lars F. Westblade ◽  
Matthew S. Simon ◽  
Blaine A. Mathison ◽  
Laura A. Kirkman

ABSTRACT Babesia microti , a zoonotic intraerythrocytic parasite, is the primary etiological agent of human babesiosis in the United States. Human infections range from subclinical illness to severe disease resulting in death, with symptoms being related to host immune status. Despite advances in our understanding and management of B. microti , the incidence of infection in the United States has increased. Therefore, research focused on eradicating disease and optimizing clinical management is essential. Here we review this remarkable organism, with emphasis on the clinical, diagnostic, and therapeutic aspects of human disease.


2000 ◽  
Vol 38 (12) ◽  
pp. 4511-4516 ◽  
Author(s):  
Atsuko Saito-Ito ◽  
Masayoshi Tsuji ◽  
Qiang Wei ◽  
Shenyi He ◽  
Toshimitsu Matsui ◽  
...  

We have isolated piroplasms from a patient who developed the first case of human babesiosis in Japan by using NOD/shi-scidmice whose circulating erythrocytes (RBCs) had been replaced with human RBCs (hu-RBC-SCID mice). Following inoculation of the patient's blood specimen into hu-RBC-SCID mice, parasites proliferated within the human RBCs in the mice, resulting in a high level of parasitemia. Parasite DNA was prepared from blood samples of the patient and the mice, and the nuclear small-subunit rRNA gene (rDNA) was amplified and sequenced. Both DNA samples gave rise to identical sequences which showed the highest degree of homology (99.2%) with the Babesia microti rDNA. Because the patient had received a blood transfusion before the onset of babesiosis, we investigated the eight donors who were involved. Their archived blood samples were analyzed for specific antibody and parasite DNA; only a single donor was found to be positive by both tests, and the parasite rDNA sequence from the donor coincided with that derived from the patient. The donor's serum exhibited a high antibody titer against the isolate from the patient, whereas it exhibited only a weak cross-reaction against B. microti strains isolated in the United States. We conclude that the first Japanese babesiosis case occurred due to a blood transfusion and that the etiological agent is an indigenous Japanese parasite which may be a geographical variant of B. microti. Our results also demonstrated the usefulness of hu-RBC-SCID mice for isolation of parasites from humans and for maintenance of the parasite infectivity for human RBCs.


2016 ◽  
Vol 82 (22) ◽  
pp. 6624-6632 ◽  
Author(s):  
Aya Zamoto-Niikura ◽  
Shigeru Morikawa ◽  
Ken-Ichi Hanaki ◽  
Patricia J. Holman ◽  
Chiaki Ishihara

ABSTRACTThe U.S. lineage, one of the major clades in theBabesia microtigroup, is known as a causal agent of human babesiosis mostly in the northeastern and upper midwestern United States. This lineage, however, also is distributed throughout the temperate zone of Eurasia with several reported human cases, although convincing evidence of the identity of the specific vector(s) in this area is lacking. Here, the goal was to demonstrate the presence of infectious parasites directly in salivary glands ofIxodes persulcatus, from which U.S. lineage genetic sequences have been detected in Asia, and to molecularly characterize the isolates. Five PCR-positive specimens were individually inoculated into hamsters, resulting in infections in four; consequently, four strains were newly established. Molecular characterization, including 18S rRNA, β-tubulin, andCCT7gene sequences, as well as Western blot analysis and indirect fluorescent antibody assay, revealed that all four strains were identical to each other and to the U.S. lineage strains isolated from rodents captured in Japan. The 18S rRNA gene sequence from the isolates was identical to those fromI. persulcatusin Russia and China, but the genetic and antigenic profiles of the Japanese parasites differ from those in the United States and Europe. Together with previous epidemiological and transmission studies, we conclude thatI. persulcatusis likely the principal vector for theB. microtiU.S. lineage in Japan and presumably in northeastern Eurasia.IMPORTANCEThe major cause of human babesiosis, the tick-borne blood parasiteBabesia microti, U.S. lineage, is widely distributed in the temperate Northern Hemisphere. However, the specific tick vector(s) remains unidentified in Eurasia, where there are people with antibodies to theB. microtiU.S. lineage and cases of human babesiosis. In this study, the first isolation ofB. microtiU.S. lineage fromIxodes persulcatusticks, a principal vector for many tick-borne diseases, is described in Japan. Limited antigenic cross-reaction was found between the Japan and United States isolates. Thus, current serological tests based on U.S. isolates may underestimateB. microtioccurrence outside the United States. This study and previous studies indicate thatI. persulcatusis part of theB. microtiU.S. lineage life cycle in Japan and, presumably, northeastern Eurasia. This report will be important for public health, especially since infection may occur through transfusion, and also to researchers in the field of parasitology.


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